Practical Chemical Synthesis of Atypical Ubiquitin Chains by Using an Isopeptide-Linked Ub Isomer - Tang, Shan; Liang, Lu-Jun; Si, Yan-Yan; Gao, Shuai; Wang, Jia-Xing; Liang, Jun; Mei, Ziqing; Zheng, Ji-Shen; Liu, Lei (Angewandte Chemie International Edition, 2017)

Scalable synthesis of γ-thiolysine starting from lysine and a side by side comparison with δ-thiolysine in non-enzymatic ubiquitination - Merkx, Remco; de Bruin, Gerjan; Kruithof, Art; van den Bergh, Toine; Snip, Erwin; Lutz, Martin; El Oualid, Farid; Ovaa, Huib (Chemical Science, 2013)

Practical Chemical Synthesis of Atypical Ubiquitin Chains by Using an Isopeptide-Linked Ub Isomer - Tang, Shan; Liang, Lu-Jun; Si, Yan-Yan; Gao, Shuai; Wang, Jia-Xing; Liang, Jun; Mei, Ziqing; Zheng, Ji-Shen; Liu, Lei (Angewandte Chemie International Edition, 2017)

Scalable synthesis of γ-thiolysine starting from lysine and a side by side comparison with δ-thiolysine in non-enzymatic ubiquitination - Merkx, Remco; de Bruin, Gerjan; Kruithof, Art; van den Bergh, Toine; Snip, Erwin; Lutz, Martin; El Oualid, Farid; Ovaa, Huib (Chemical Science, 2013)

Chemical Synthesis of Ubiquitin, Ubiquitin-Based Probes, and Diubiquitin - El Oualid, Farid; Merkx, Remco; Ekkebus, Reggy; Hameed, Dharjath S.; Smit, Judith J.; de Jong, Annemieke; Hilkmann, Henk; Sixma, Titia K.; Ovaa, Huib (Angewandte Chemie International Edition, 2010)

Chemical Synthesis of Ubiquitin, Ubiquitin-Based Probes, and Diubiquitin - El Oualid, Farid; Merkx, Remco; Ekkebus, Reggy; Hameed, Dharjath S.; Smit, Judith J.; de Jong, Annemieke; Hilkmann, Henk; Sixma, Titia K.; Ovaa, Huib (Angewandte Chemie International Edition, 2010)

Total Chemical Synthesis of Di-ubiquitin Chains - Kumar, K. S. Ajish; Spasser, Liat; Erlich, Lesly A.; Bavikar, Sudhir N.; Brik, Ashraf (Angewandte Chemie, 2010)

Total Chemical Synthesis of Di-ubiquitin Chains - Kumar, K. S. Ajish; Spasser, Liat; Erlich, Lesly A.; Bavikar, Sudhir N.; Brik, Ashraf (Angewandte Chemie, 2010)

Total Chemical Synthesis of Di-ubiquitin Chains - Kumar, K. S. Ajish; Spasser, Liat; Erlich, Lesly A.; Bavikar, Sudhir N.; Brik, Ashraf (Angewandte Chemie International Edition, 2010)

Total Chemical Synthesis of Di-ubiquitin Chains - Kumar, K. S. Ajish; Spasser, Liat; Erlich, Lesly A.; Bavikar, Sudhir N.; Brik, Ashraf (Angewandte Chemie International Edition, 2010)

Synthesis of K48-linked diubiquitin using dual native chemical ligation at lysine - Yang, Renliang; Pasunooti, Kalyan Kumar; Li, Fupeng; Liu, Xue-Wei; Liu, Chuan-Fa (Chemical Communications, 2010)

Synthesis of K48-linked diubiquitin using dual native chemical ligation at lysine - Yang, Renliang; Pasunooti, Kalyan Kumar; Li, Fupeng; Liu, Xue-Wei; Liu, Chuan-Fa (Chemical Communications, 2010)

Chemical Synthesis of Phosphorylated Ubiquitin and Diubiquitin Exposes Positional Sensitivities of E1-E2 Enzymes and Deubiquitinases - Bondalapati, Somasekhar; Mansour, Wissam; Nakasone, Mark A.; Maity, Suman Kumar; Glickman, Michael H.; Brik, Ashraf (Chemistry - A European Journal, 2015)

Chemical Synthesis of Phosphorylated Ubiquitin and Diubiquitin Exposes Positional Sensitivities of E1-E2 Enzymes and Deubiquitinases - Bondalapati, Somasekhar; Mansour, Wissam; Nakasone, Mark A.; Maity, Suman Kumar; Glickman, Michael H.; Brik, Ashraf (Chemistry - A European Journal, 2015)

Highly efficient one-pot ligation and desulfurization - Moyal, Tal; Hemantha, Hosahalli P.; Siman, Peter; Refua, Maya; Brik, Ashraf (Chemical Science, 2013)

Highly efficient one-pot ligation and desulfurization - Moyal, Tal; Hemantha, Hosahalli P.; Siman, Peter; Refua, Maya; Brik, Ashraf (Chemical Science, 2013)

One-pot ligation strategy for atypical ubiquitin chains synthesis by using the trifluoroacetamidomethyl-protected isopeptide-linked Ub (Tfacm-isoUb) unit - Si, Yan-Yan; Liang, Lu-Jun; Tang, Shan; Qi, Yun-Kun; Huang, Yong; Zheng, Ji-Shen (Tetrahedron Letters, 2018)

One-pot ligation strategy for atypical ubiquitin chains synthesis by using the trifluoroacetamidomethyl-protected isopeptide-linked Ub (Tfacm-isoUb) unit - Si, Yan-Yan; Liang, Lu-Jun; Tang, Shan; Qi, Yun-Kun; Huang, Yong; Zheng, Ji-Shen (Tetrahedron Letters, 2018)

Native chemical ubiquitination using a genetically incorporated azidonorleucine - Yang, Renliang; Bi, Xiaobao; Li, Fupeng; Cao, Yuan; Liu, Chuan-Fa (Chem. Commun., 2014)
A genetically incorporated azidonorleucine serves as a lysine precursor for site-selective ubiquitination of a recombinant protein through an auxiliary group. , A robust chemical ubiquitination method was developed. The method employed a genetically incorporated azidonorleucine as an orthogonal lysine precursor for the installation of a Gly residue bearing an Nα-auxiliary which mediated the ligation between ubiquitin(1–75)-thioester and the target protein. To demonstrate our methodology, a model protein, K48-linked diubiquitin, was synthesized with an overall yield of 35%.

Native chemical ubiquitination using a genetically incorporated azidonorleucine - Yang, Renliang; Bi, Xiaobao; Li, Fupeng; Cao, Yuan; Liu, Chuan-Fa (Chem. Commun., 2014)
A genetically incorporated azidonorleucine serves as a lysine precursor for site-selective ubiquitination of a recombinant protein through an auxiliary group. , A robust chemical ubiquitination method was developed. The method employed a genetically incorporated azidonorleucine as an orthogonal lysine precursor for the installation of a Gly residue bearing an Nα-auxiliary which mediated the ligation between ubiquitin(1–75)-thioester and the target protein. To demonstrate our methodology, a model protein, K48-linked diubiquitin, was synthesized with an overall yield of 35%.

Native chemical ligation at methionine bioisostere norleucine allows for N-terminal chemical protein ligation - Xin, Bo-Tao; van Tol, Bianca D. M.; Ovaa, Huib; Geurink, Paul P. (Organic & Biomolecular Chemistry, 2018)
γ-Thionorleucine is synthesized and used for N-terminal chemical protein modification by native chemical ligation–desulfurization to prepare linear diubiquitin. , The development of γ-thionorleucine (ThioNle) as a handle for native chemical ligation–desulfurization is reported here. ThioNle is a new addition to the expanding thiolated amino acid toolbox and serves as a methionine substitute in NCL with the advantage that it lacks the undesirable oxidation-prone thioether moiety. Its usefulness for N-terminal ubiquitination is demonstrated by efficient preparation of fully synthetic linear diubiquitin with preserved protein folding compared to the expressed material. Interestingly, gel-based deubiquitinating assays revealed that the methionine to norleucine substitution did affect diubiquitin cleavage, which may indicate a more profound role for methionine in the interaction between ubiquitin and the deubiquitinating enzymes than has been known so far.

Native chemical ligation at methionine bioisostere norleucine allows for N-terminal chemical protein ligation - Xin, Bo-Tao; van Tol, Bianca D. M.; Ovaa, Huib; Geurink, Paul P. (Organic & Biomolecular Chemistry, 2018)
γ-Thionorleucine is synthesized and used for N-terminal chemical protein modification by native chemical ligation–desulfurization to prepare linear diubiquitin. , The development of γ-thionorleucine (ThioNle) as a handle for native chemical ligation–desulfurization is reported here. ThioNle is a new addition to the expanding thiolated amino acid toolbox and serves as a methionine substitute in NCL with the advantage that it lacks the undesirable oxidation-prone thioether moiety. Its usefulness for N-terminal ubiquitination is demonstrated by efficient preparation of fully synthetic linear diubiquitin with preserved protein folding compared to the expressed material. Interestingly, gel-based deubiquitinating assays revealed that the methionine to norleucine substitution did affect diubiquitin cleavage, which may indicate a more profound role for methionine in the interaction between ubiquitin and the deubiquitinating enzymes than has been known so far.

A Highly Efficient Synthesis of Polyubiquitin Chains - Qu, Qian; Pan, Man; Gao, Shuai; Zheng, Qing-Yun; Yu, Yuan-Yuan; Su, Jia-Can; Li, Xiang; Hu, Hong-Gang (Advanced Science, 2018)

A Highly Efficient Synthesis of Polyubiquitin Chains - Qu, Qian; Pan, Man; Gao, Shuai; Zheng, Qing-Yun; Yu, Yuan-Yuan; Su, Jia-Can; Li, Xiang; Hu, Hong-Gang (Advanced Science, 2018)

N, P, and S Codoped Graphene-Like Carbon Nanosheets for Ultrafast Uranium (VI) Capture with High Capacity - Chen, Zhe; Chen, Wanying; Jia, Dashuang; Liu, Yang; Zhang, Anrui; Wen, Tao; Liu, Jian; Ai, Yuejie; Song, Weiguo; Wang, Xiangke (Advanced Science, 2018)

N, P, and S Codoped Graphene-Like Carbon Nanosheets for Ultrafast Uranium (VI) Capture with High Capacity - Chen, Zhe; Chen, Wanying; Jia, Dashuang; Liu, Yang; Zhang, Anrui; Wen, Tao; Liu, Jian; Ai, Yuejie; Song, Weiguo; Wang, Xiangke (Advanced Science, 2018)

Independent Band Modulation in 2D van der Waals Heterostructures via a Novel Device Architecture - Guo, Zhongxun; Chen, Yan; Zhang, Heng; Wang, Jianlu; Hu, Weida; Ding, Shijin; Zhang, David Wei; Zhou, Peng; Bao, Wenzhong (Advanced Science, 2018)

Independent Band Modulation in 2D van der Waals Heterostructures via a Novel Device Architecture - Guo, Zhongxun; Chen, Yan; Zhang, Heng; Wang, Jianlu; Hu, Weida; Ding, Shijin; Zhang, David Wei; Zhou, Peng; Bao, Wenzhong (Advanced Science, 2018)

Self-Sensing Paper Actuators Based on Graphite-Carbon Nanotube Hybrid Films - Amjadi, Morteza; Sitti, Metin (Advanced Science, 2018)

Self-Sensing Paper Actuators Based on Graphite-Carbon Nanotube Hybrid Films - Amjadi, Morteza; Sitti, Metin (Advanced Science, 2018)

Multimeric Amphipathic α-Helical Sequences for Rapid and Efficient Intracellular Protein Transport at Nanomolar Concentrations - Oh, Jae Hoon; Chong, Seung-Eun; Nam, Sohee; Hyun, Soonsil; Choi, Sejong; Gye, Hyojun; Jang, Sangmok; Jang, Joomyung; Hwang, Sung Won; Yu, Jaehoon; Lee, Yan (Advanced Science, 2018)

Multimeric Amphipathic α-Helical Sequences for Rapid and Efficient Intracellular Protein Transport at Nanomolar Concentrations - Oh, Jae Hoon; Chong, Seung-Eun; Nam, Sohee; Hyun, Soonsil; Choi, Sejong; Gye, Hyojun; Jang, Sangmok; Jang, Joomyung; Hwang, Sung Won; Yu, Jaehoon; Lee, Yan (Advanced Science, 2018)

Chemical Synthesis of K48-Linked Diubiquitin by Incorporation of a Lysine-Linked Auxiliary Handle - Xie, Rui-Long; Xu, Ling; Li, Jia-Bin; Chu, Guo-Chao; Wang, Tao; Huang, Yi-Chao; Li, Yi-Ming (European Journal of Organic Chemistry, 2016)

Chemical Synthesis of K48-Linked Diubiquitin by Incorporation of a Lysine-Linked Auxiliary Handle - Xie, Rui-Long; Xu, Ling; Li, Jia-Bin; Chu, Guo-Chao; Wang, Tao; Huang, Yi-Chao; Li, Yi-Ming (European Journal of Organic Chemistry, 2016)

Chemical Synthesis of Structurally Defined Phosphorylated Ubiquitins Suggests Impaired Parkin Activation by Phosphorylated Ubiquitins with a Non-Phosphorylated Distal Unit - Pan, Man; Zheng, Qingyun; Gao, Shuai; Qu, Qian; Yu, Yuanyuan; Wu, Ming; Lan, Huan; Li, Yulei; Liu, Sanling; Li, Jiabin; Sun, Demeng; Lu, Lining; et al. (CCS Chemistry, 2019)

Chemical Synthesis of Structurally Defined Phosphorylated Ubiquitins Suggests Impaired Parkin Activation by Phosphorylated Ubiquitins with a Non-Phosphorylated Distal Unit - Pan, Man; Zheng, Qingyun; Gao, Shuai; Qu, Qian; Yu, Yuanyuan; Wu, Ming; Lan, Huan; Li, Yulei; Liu, Sanling; Li, Jiabin; Sun, Demeng; Lu, Lining; et al. (CCS Chemistry, 2019)

Semisynthesis and initial characterization of sortase A mutants containing selenocysteine and homocysteine - Schmohl, Lena; Wagner, Felix Roman; Schümann, Michael; Krause, Eberhard; Schwarzer, Dirk (Bioorganic & Medicinal Chemistry, 2015)

Total Chemical Synthesis of the Enzyme Sortase A ΔN59 with Full Catalytic Activity - Deng, Fang-Kun; Zhang, Liang; Wang, Ya-Ting; Schneewind, Olaf; Kent, Stephen B. H. (Angewandte Chemie International Edition, 2014)

A Threonine‐Forming Oxazetidine Amino Acid for the Chemical Synthesis of Proteins through KAHA Ligation - Baldauf, Simon; Schauenburg, Dominik; Bode, Jeffrey W. (Angewandte Chemie International Edition, 2019)

Native N-glycopeptide thioester synthesis through N→S acyl transfer - Premdjee, Bhavesh; Adams, Anna L.; Macmillan, Derek (Bioorganic & Medicinal Chemistry Letters, 2011)

Chemical Synthesis of the β-Subunit of Human Luteinizing (hLH) and Chorionic Gonadotropin (hCG) Glycoprotein Hormones - Fernández-Tejada, Alberto; Vadola, Paul A.; Danishefsky, Samuel J. (Journal of the American Chemical Society, 2014)

Accessing human selenoproteins through chemical protein synthesis - Dery, L.; Reddy, P. Sai; Dery, S.; Mousa, R.; Ktorza, O.; Talhami, A.; Metanis, N. (Chemical Science, 2017)
The human body contains 25 selenoproteins, but challenges in their preparations have prevented biological characterizations thus far. Here we report the first total chemical syntheses of two human selenoproteins, selenoprotein M (SELM) and selenoprotein W (SELW). , The human body contains 25 selenoproteins, which contain in their sequence the twenty-first encoded amino acid, selenocysteine. About a dozen of these proteins remain functionally uncharacterized or poorly studied. Challenges in accessing these selenoproteins using traditional recombinant expressions have prevented biological characterization thus far. Chemical protein synthesis has the potential to overcome these hurdles. Here we report the first total chemical syntheses of two human selenoproteins, selenoprotein M (SELM) and selenoprotein W (SELW). The synthesis of the more challenging protein SELM was enabled using recent advances in the field of selenocysteine chemistry. This approach allows the preparation of selenoproteins in milligram quantities and in homogenous form, which should open new horizons for future studies to pursue a fuller biological understanding of their role in health and disease.

Semisynthesis and initial characterization of sortase A mutants containing selenocysteine and homocysteine - Schmohl, Lena; Wagner, Felix Roman; Schümann, Michael; Krause, Eberhard; Schwarzer, Dirk (Bioorganic & Medicinal Chemistry, 2015)

Total Chemical Synthesis of the Enzyme Sortase A ΔN59 with Full Catalytic Activity - Deng, Fang-Kun; Zhang, Liang; Wang, Ya-Ting; Schneewind, Olaf; Kent, Stephen B. H. (Angewandte Chemie International Edition, 2014)

Enabling N-to-C Ser/Thr Ligation for Convergent Protein Synthesis via Combining Chemical Ligation Approaches - Lee, Chi Lung; Liu, Han; Wong, Clarence T. T.; Chow, Hoi Yee; Li, Xuechen (Journal of the American Chemical Society, 2016)

A Threonine‐Forming Oxazetidine Amino Acid for the Chemical Synthesis of Proteins through KAHA Ligation - Baldauf, Simon; Schauenburg, Dominik; Bode, Jeffrey W. (Angewandte Chemie International Edition, 2019)

Synthesis of misfolded glycoprotein dimers through native chemical ligation of a dimeric peptide thioester - Izumi, Masayuki; Komaki, Shinji; Okamoto, Ryo; Seko, Akira; Takeda, Yoichi; Ito, Yukishige; Kajihara, Yasuhiro (Organic & Biomolecular Chemistry, 2016)
Misfolded glycoprotein dimers were synthesized through double native chemical ligation between a dimeric peptide-α-thioester, glycopeptide, and non-glycosylated peptide. , Glycoprotein quality control processes are very important for an efficient production of glycoproteins and for avoiding the accumulation of unwanted toxic species in cells. These complex processes consist of multiple enzymes and chaperones such as UGGT, calnexin/calreticulin, and glucosidase II. We designed and synthesized monomeric and dimeric misfolded glycoprotein probes. Synthetic homogeneous monomeric glycoproteins proved to be useful substrates for kinetic analyses of the folding sensor enzyme UGGT. For a concise synthesis of a bismaleimide-linked dimer, we examined double native chemical ligation (dNCL) of a dimeric peptide-α-thioester. The dNCL to two equivalents of glycopeptides gave a homodimer. The dNCL to a 1 : 1 mixture of a glycopeptide and a non-glycosylated peptide gave all the three possible ligation products consisting of two homodimers and a heterodimer. Both the homodimer bearing two Man 9 GlcNAc 2 (M9) oligosaccharides and the heterodimer bearing one M9 oligosaccharide were found to be good substrates of UGGT.

Native N-glycopeptide thioester synthesis through N→S acyl transfer - Premdjee, Bhavesh; Adams, Anna L.; Macmillan, Derek (Bioorganic & Medicinal Chemistry Letters, 2011)

Convergent Chemical Synthesis of Proteins by Ligation of Peptide Hydrazides - Fang, Ge-Min; Wang, Jia-Xing; Liu, Lei (Angewandte Chemie, 2012)

Chemically synthesized human survivin does not inhibit caspase-3 - Li, Changqing; Wu, Zhibin; Liu, Min; Pazgier, Marzena; Lu, Wuyuan (Protein Science, 2008)

Chemical Synthesis of the β-Subunit of Human Luteinizing (hLH) and Chorionic Gonadotropin (hCG) Glycoprotein Hormones - Fernández-Tejada, Alberto; Vadola, Paul A.; Danishefsky, Samuel J. (Journal of the American Chemical Society, 2014)

Triple Function of 4-Mercaptophenylacetic Acid Promotes One-Pot Multiple Peptide Ligation - Kamo, Naoki; Hayashi, Gosuke; Okamoto, Akimitsu (Angewandte Chemie International Edition, 2018)

Accessing human selenoproteins through chemical protein synthesis - Dery, L.; Reddy, P. Sai; Dery, S.; Mousa, R.; Ktorza, O.; Talhami, A.; Metanis, N. (Chemical Science, 2017)
The human body contains 25 selenoproteins, but challenges in their preparations have prevented biological characterizations thus far. Here we report the first total chemical syntheses of two human selenoproteins, selenoprotein M (SELM) and selenoprotein W (SELW). , The human body contains 25 selenoproteins, which contain in their sequence the twenty-first encoded amino acid, selenocysteine. About a dozen of these proteins remain functionally uncharacterized or poorly studied. Challenges in accessing these selenoproteins using traditional recombinant expressions have prevented biological characterization thus far. Chemical protein synthesis has the potential to overcome these hurdles. Here we report the first total chemical syntheses of two human selenoproteins, selenoprotein M (SELM) and selenoprotein W (SELW). The synthesis of the more challenging protein SELM was enabled using recent advances in the field of selenocysteine chemistry. This approach allows the preparation of selenoproteins in milligram quantities and in homogenous form, which should open new horizons for future studies to pursue a fuller biological understanding of their role in health and disease.

Chemistry as an Expanding Resource in Protein Science: Fully Synthetic and Fully Active Human Parathyroid Hormone-Related Protein (1 - 141) - Li, Jianfeng; Dong, Suwei; Townsend, Steven D.; Dean, Thomas; Gardella, Thomas J.; Danishefsky, Samuel J. (Angewandte Chemie, 2012)

Site-Specific Three-Color Labeling of α-Synuclein via Conjugation to Uniquely Reactive Cysteines during Assembly by Native Chemical Ligation - Lee, Taehyung C.; Moran, Crystal R.; Cistrone, Philip A.; Dawson, Philip E.; Deniz, Ashok A. (Cell Chemical Biology, 2018)

Enabling N-to-C Ser/Thr Ligation for Convergent Protein Synthesis via Combining Chemical Ligation Approaches - Lee, Chi Lung; Liu, Han; Wong, Clarence T. T.; Chow, Hoi Yee; Li, Xuechen (Journal of the American Chemical Society, 2016)

Synthesis of misfolded glycoprotein dimers through native chemical ligation of a dimeric peptide thioester - Izumi, Masayuki; Komaki, Shinji; Okamoto, Ryo; Seko, Akira; Takeda, Yoichi; Ito, Yukishige; Kajihara, Yasuhiro (Organic & Biomolecular Chemistry, 2016)
Misfolded glycoprotein dimers were synthesized through double native chemical ligation between a dimeric peptide-α-thioester, glycopeptide, and non-glycosylated peptide. , Glycoprotein quality control processes are very important for an efficient production of glycoproteins and for avoiding the accumulation of unwanted toxic species in cells. These complex processes consist of multiple enzymes and chaperones such as UGGT, calnexin/calreticulin, and glucosidase II. We designed and synthesized monomeric and dimeric misfolded glycoprotein probes. Synthetic homogeneous monomeric glycoproteins proved to be useful substrates for kinetic analyses of the folding sensor enzyme UGGT. For a concise synthesis of a bismaleimide-linked dimer, we examined double native chemical ligation (dNCL) of a dimeric peptide-α-thioester. The dNCL to two equivalents of glycopeptides gave a homodimer. The dNCL to a 1 : 1 mixture of a glycopeptide and a non-glycosylated peptide gave all the three possible ligation products consisting of two homodimers and a heterodimer. Both the homodimer bearing two Man 9 GlcNAc 2 (M9) oligosaccharides and the heterodimer bearing one M9 oligosaccharide were found to be good substrates of UGGT.

Convergent Chemical Synthesis of Proteins by Ligation of Peptide Hydrazides - Fang, Ge-Min; Wang, Jia-Xing; Liu, Lei (Angewandte Chemie, 2012)

Chemically synthesized human survivin does not inhibit caspase-3 - Li, Changqing; Wu, Zhibin; Liu, Min; Pazgier, Marzena; Lu, Wuyuan (Protein Science, 2008)

Characterization of Semisynthetic and Naturally N α -Acetylated α-Synuclein in Vitro and in Intact Cells: IMPLICATIONS FOR AGGREGATION AND CELLULAR PROPERTIES OF α-SYNUCLEIN - Fauvet, Bruno; Fares, Mohamed-Bilal; Samuel, Filsy; Dikiy, Igor; Tandon, Anurag; Eliezer, David; Lashuel, Hilal A. (Journal of Biological Chemistry, 2012)

Triple Function of 4-Mercaptophenylacetic Acid Promotes One-Pot Multiple Peptide Ligation - Kamo, Naoki; Hayashi, Gosuke; Okamoto, Akimitsu (Angewandte Chemie International Edition, 2018)

Chemistry as an Expanding Resource in Protein Science: Fully Synthetic and Fully Active Human Parathyroid Hormone-Related Protein (1 - 141) - Li, Jianfeng; Dong, Suwei; Townsend, Steven D.; Dean, Thomas; Gardella, Thomas J.; Danishefsky, Samuel J. (Angewandte Chemie, 2012)

Site-Specific Three-Color Labeling of α-Synuclein via Conjugation to Uniquely Reactive Cysteines during Assembly by Native Chemical Ligation - Lee, Taehyung C.; Moran, Crystal R.; Cistrone, Philip A.; Dawson, Philip E.; Deniz, Ashok A. (Cell Chemical Biology, 2018)

Characterization of Semisynthetic and Naturally N α -Acetylated α-Synuclein in Vitro and in Intact Cells: IMPLICATIONS FOR AGGREGATION AND CELLULAR PROPERTIES OF α-SYNUCLEIN - Fauvet, Bruno; Fares, Mohamed-Bilal; Samuel, Filsy; Dikiy, Igor; Tandon, Anurag; Eliezer, David; Lashuel, Hilal A. (Journal of Biological Chemistry, 2012)

Native Chemical Ligation of Thioamide-Containing Peptides: Development and Application to the Synthesis of Labeled α-Synuclein for Misfolding Studies - Batjargal, Solongo; Wang, Yanxin J.; Goldberg, Jacob M.; Wissner, Rebecca F.; Petersson, E. James (Journal of the American Chemical Society, 2012)

Hybrid phase ligation for efficient synthesis of histone proteins - Yu, Ruixuan R.; Mahto, Santosh K.; Justus, Kurt; Alexander, Mallory M.; Howard, Cecil J.; Ottesen, Jennifer J. (Organic & Biomolecular Chemistry, 2016)
We report a hybrid solid-solution phase ligation approach that combines the efficiency of solid phase ligation with solution phase ligation in the total synthesis of modified histone proteins. , We introduce a hybrid solid-solution phase ligation approach that combines the efficiency of solid phase ligation with solution phase ligation in the total synthesis of modified histone proteins. A two linker strategy allows analysis throughout work on the solid phase and maximizes yields through cleavage at an external Rink, while an internal HMBA linker allows the native carboxyl terminus for any protein sequence. We demonstrate this approach for two histone proteins: triple-acetylated H4-K5ac, K12ac, K91ac and CENP-A-K124ac.

Native Chemical Ligation of Thioamide-Containing Peptides: Development and Application to the Synthesis of Labeled α-Synuclein for Misfolding Studies - Batjargal, Solongo; Wang, Yanxin J.; Goldberg, Jacob M.; Wissner, Rebecca F.; Petersson, E. James (Journal of the American Chemical Society, 2012)

O-GlcNAc modification blocks the aggregation and toxicity of the protein α-synuclein associated with Parkinson's disease - Marotta, Nicholas P.; Lin, Yu Hsuan; Lewis, Yuka E.; Ambroso, Mark R.; Zaro, Balyn W.; Roth, Maxwell T.; Arnold, Don B.; Langen, Ralf; Pratt, Matthew R. (Nature Chemistry, 2015)

One-pot total chemical synthesis of human α-synuclein - Fauvet, Bruno; Butterfield, Sara M.; Fuks, Jonas; Brik, Ashraf; Lashuel, Hilal A. (Chemical Communications, 2013)

Hybrid phase ligation for efficient synthesis of histone proteins - Yu, Ruixuan R.; Mahto, Santosh K.; Justus, Kurt; Alexander, Mallory M.; Howard, Cecil J.; Ottesen, Jennifer J. (Organic & Biomolecular Chemistry, 2016)
We report a hybrid solid-solution phase ligation approach that combines the efficiency of solid phase ligation with solution phase ligation in the total synthesis of modified histone proteins. , We introduce a hybrid solid-solution phase ligation approach that combines the efficiency of solid phase ligation with solution phase ligation in the total synthesis of modified histone proteins. A two linker strategy allows analysis throughout work on the solid phase and maximizes yields through cleavage at an external Rink, while an internal HMBA linker allows the native carboxyl terminus for any protein sequence. We demonstrate this approach for two histone proteins: triple-acetylated H4-K5ac, K12ac, K91ac and CENP-A-K124ac.

O-GlcNAc modification blocks the aggregation and toxicity of the protein α-synuclein associated with Parkinson's disease - Marotta, Nicholas P.; Lin, Yu Hsuan; Lewis, Yuka E.; Ambroso, Mark R.; Zaro, Balyn W.; Roth, Maxwell T.; Arnold, Don B.; Langen, Ralf; Pratt, Matthew R. (Nature Chemistry, 2015)

Elucidating the Role of Site-Specific Nitration of α-Synuclein in the Pathogenesis of Parkinson’s Disease via Protein Semisynthesis and Mutagenesis - Burai, Ritwik; Ait-Bouziad, Nadine; Chiki, Anass; Lashuel, Hilal A. (Journal of the American Chemical Society, 2015)

Semi‐synthesis of murine prion protein by native chemical ligation and chemical activation for preparation of polypeptide‐ α ‐thioester - Shi, Lei; Chen, Huai; Zhang, Si‐Yu; Chu, Ting‐Ting; Zhao, Yu‐Fen; Chen, Yong‐Xiang; Li, Yan‐Mei (Journal of Peptide Science, 2017)

One-pot total chemical synthesis of human α-synuclein - Fauvet, Bruno; Butterfield, Sara M.; Fuks, Jonas; Brik, Ashraf; Lashuel, Hilal A. (Chemical Communications, 2013)

O-GlcNAc modification inhibits the calpain-mediated cleavage of α-synuclein - Levine, Paul M.; De Leon, Cesar A.; Galesic, Ana; Balana, Aaron; Marotta, Nicholas P.; Lewis, Yuka E.; Pratt, Matthew R. (Bioorganic & Medicinal Chemistry, 2017)

Elucidating the Role of Site-Specific Nitration of α-Synuclein in the Pathogenesis of Parkinson’s Disease via Protein Semisynthesis and Mutagenesis - Burai, Ritwik; Ait-Bouziad, Nadine; Chiki, Anass; Lashuel, Hilal A. (Journal of the American Chemical Society, 2015)

Comparison of strategies for non-perturbing labeling of α-synuclein to study amyloidogenesis - Haney, Conor M.; Wissner, Rebecca F.; Warner, John B.; Wang, Yanxin J.; Ferrie, John J.; J. Covell, Dustin; Karpowicz, Richard J.; Lee, Virginia M.-Y.; James Petersson, E. (Organic & Biomolecular Chemistry, 2016)
Complementary methods for site-specific protein labeling are used to introduce fluorescent probes into the Parkinson's Disease protein α-synuclein to examine fibril formation and cellular uptake. , Characterization of the amyloidogenic Parkinson's disease protein α-synuclein (αS) has proven difficult due to its structural plasticity. Here, we present a number of complementary methods to site-specifically introduce fluorescent probes to examine αS fibril formation and cellular uptake. By using various combinations of conventional Cys modification, amber codon suppression, transferase mediated N-terminal modification, and native chemical ligation, several variants of singly- and doubly-labeled αS were produced. We validated the nonperturbative nature of the label by a combination of in vitro aggregation kinetics measurements and imaging of the resulting fibrils. The labeled αS can then be used to monitor conformational changes during fibril formation or cellular uptake of αS fibrils in models of disease propagation.

Semi‐synthesis of murine prion protein by native chemical ligation and chemical activation for preparation of polypeptide‐ α ‐thioester - Shi, Lei; Chen, Huai; Zhang, Si‐Yu; Chu, Ting‐Ting; Zhao, Yu‐Fen; Chen, Yong‐Xiang; Li, Yan‐Mei (Journal of Peptide Science, 2017)

O-GlcNAc modification inhibits the calpain-mediated cleavage of α-synuclein - Levine, Paul M.; De Leon, Cesar A.; Galesic, Ana; Balana, Aaron; Marotta, Nicholas P.; Lewis, Yuka E.; Pratt, Matthew R. (Bioorganic & Medicinal Chemistry, 2017)

α-Synuclein O-GlcNAcylation alters aggregation and toxicity, revealing certain residues as potential inhibitors of Parkinson’s disease - Levine, Paul M.; Galesic, Ana; Balana, Aaron T.; Mahul-Mellier, Anne-Laure; Navarro, Mariana X.; De Leon, Cesar A.; Lashuel, Hilal A.; Pratt, Matthew R. (Proceedings of the National Academy of Sciences, 2019)
A compelling link is emerging between the posttranslational modification O-GlcNAc and protein aggregation. A prime example is α-synuclein, which forms toxic aggregates that are associated with neurodegeneration in Parkinson’s and related diseases. α-Synuclein has been shown to be O-GlcNAcylated at nine different positions in in vivo proteomics experiments from mouse and human tissues. This raises the possibility that O-GlcNAc may alter the aggregation of this protein and could be both an important biological mediator of neurodegeneration and also a therapeutic target. Here, we expand upon our previous research in this area through the chemical synthesis of six site-specifically O-GlcNAcylated variants of α-synuclein. We then use a variety of biochemical experiments to show that O-GlcNAc in general inhibits the aggregation of α-synuclein but can also alter the structure of α-synuclein aggregates in site-specific ways. Additionally, an α-synuclein protein bearing three O-GlcNAc modifications can inhibit the aggregation of unmodified protein. Primary cell culture experiments also show that several of the O-GlcNAc sites inhibit the toxicity of extracellular α-synuclein fibers that are likely culprits in the spread of Parkinson’s disease. We also demonstrate that O-GlcNAcylation can inhibit the aggregation of an aggressive mutant of α-synuclein, indicating that therapies currently in development that increase this modification might be applied in animal models that rely on this mutant. Finally, we also show that the pan-selective antibody for O-GlcNAc does not generally recognize this modification on α-synuclein, potentially explaining why it remains understudied. These results support further development of O-GlcNAcylation tools and therapeutic strategies in neurodegenerative diseases.

Comparison of strategies for non-perturbing labeling of α-synuclein to study amyloidogenesis - Haney, Conor M.; Wissner, Rebecca F.; Warner, John B.; Wang, Yanxin J.; Ferrie, John J.; J. Covell, Dustin; Karpowicz, Richard J.; Lee, Virginia M.-Y.; James Petersson, E. (Organic & Biomolecular Chemistry, 2016)
Complementary methods for site-specific protein labeling are used to introduce fluorescent probes into the Parkinson's Disease protein α-synuclein to examine fibril formation and cellular uptake. , Characterization of the amyloidogenic Parkinson's disease protein α-synuclein (αS) has proven difficult due to its structural plasticity. Here, we present a number of complementary methods to site-specifically introduce fluorescent probes to examine αS fibril formation and cellular uptake. By using various combinations of conventional Cys modification, amber codon suppression, transferase mediated N-terminal modification, and native chemical ligation, several variants of singly- and doubly-labeled αS were produced. We validated the nonperturbative nature of the label by a combination of in vitro aggregation kinetics measurements and imaging of the resulting fibrils. The labeled αS can then be used to monitor conformational changes during fibril formation or cellular uptake of αS fibrils in models of disease propagation.

α-Synuclein O-GlcNAcylation alters aggregation and toxicity, revealing certain residues as potential inhibitors of Parkinson’s disease - Levine, Paul M.; Galesic, Ana; Balana, Aaron T.; Mahul-Mellier, Anne-Laure; Navarro, Mariana X.; De Leon, Cesar A.; Lashuel, Hilal A.; Pratt, Matthew R. (Proceedings of the National Academy of Sciences, 2019)
A compelling link is emerging between the posttranslational modification O-GlcNAc and protein aggregation. A prime example is α-synuclein, which forms toxic aggregates that are associated with neurodegeneration in Parkinson’s and related diseases. α-Synuclein has been shown to be O-GlcNAcylated at nine different positions in in vivo proteomics experiments from mouse and human tissues. This raises the possibility that O-GlcNAc may alter the aggregation of this protein and could be both an important biological mediator of neurodegeneration and also a therapeutic target. Here, we expand upon our previous research in this area through the chemical synthesis of six site-specifically O-GlcNAcylated variants of α-synuclein. We then use a variety of biochemical experiments to show that O-GlcNAc in general inhibits the aggregation of α-synuclein but can also alter the structure of α-synuclein aggregates in site-specific ways. Additionally, an α-synuclein protein bearing three O-GlcNAc modifications can inhibit the aggregation of unmodified protein. Primary cell culture experiments also show that several of the O-GlcNAc sites inhibit the toxicity of extracellular α-synuclein fibers that are likely culprits in the spread of Parkinson’s disease. We also demonstrate that O-GlcNAcylation can inhibit the aggregation of an aggressive mutant of α-synuclein, indicating that therapies currently in development that increase this modification might be applied in animal models that rely on this mutant. Finally, we also show that the pan-selective antibody for O-GlcNAc does not generally recognize this modification on α-synuclein, potentially explaining why it remains understudied. These results support further development of O-GlcNAcylation tools and therapeutic strategies in neurodegenerative diseases.

α-Selective Lysine Ligation and Application in Chemical Synthesis of Interferon Gamma - Dao, Yuankun; Han, Lin; Wang, Hanxuan; Dong, Suwei (Organic Letters, 2019)

Chromatin Affinity Purification and Quantitative Mass Spectrometry Defining the Interactome of Histone Modification Patterns - Nikolov, Miroslav; Stützer, Alexandra; Mosch, Kerstin; Krasauskas, Andrius; Soeroes, Szabolcs; Stark, Holger; Urlaub, Henning; Fischle, Wolfgang (Molecular & Cellular Proteomics, 2011)

α-Selective Lysine Ligation and Application in Chemical Synthesis of Interferon Gamma - Dao, Yuankun; Han, Lin; Wang, Hanxuan; Dong, Suwei (Organic Letters, 2019)

Chromatin Affinity Purification and Quantitative Mass Spectrometry Defining the Interactome of Histone Modification Patterns - Nikolov, Miroslav; Stützer, Alexandra; Mosch, Kerstin; Krasauskas, Andrius; Soeroes, Szabolcs; Stark, Holger; Urlaub, Henning; Fischle, Wolfgang (Molecular & Cellular Proteomics, 2011)

Sequential Peptide Ligation by Combining the Cys–Pro Ester (CPE) and Thioester Methods and Its Application to the Synthesis of Histone H3 Containing a Trimethyl Lysine Residue - Kawakami, Toru; Akai, Yuichi; Fujimoto, Hisao; Kita, Chieko; Aoki, Yuko; Konishi, Takehiko; Waseda, Masazumi; Takemura, Lisa; Aimoto, Saburo (Bulletin of the Chemical Society of Japan, 2013)

Sequential Peptide Ligation by Combining the Cys–Pro Ester (CPE) and Thioester Methods and Its Application to the Synthesis of Histone H3 Containing a Trimethyl Lysine Residue - Kawakami, Toru; Akai, Yuichi; Fujimoto, Hisao; Kita, Chieko; Aoki, Yuko; Konishi, Takehiko; Waseda, Masazumi; Takemura, Lisa; Aimoto, Saburo (Bulletin of the Chemical Society of Japan, 2013)

Solid phase chemical ligation employing a rink amide linker for the synthesis of histone H2B protein - Jbara, Muhammad; Seenaiah, Mallikanti; Brik, Ashraf (Chem. Commun., 2014)

Solid phase chemical ligation employing a rink amide linker for the synthesis of histone H2B protein - Jbara, Muhammad; Seenaiah, Mallikanti; Brik, Ashraf (Chem. Commun., 2014)

One-pot native chemical ligation of peptide hydrazides enables total synthesis of modified histones - Li, Jiabin; Li, Yuanyuan; He, Qiaoqiao; Li, Yiming; Li, Haitao; Liu, Lei (Org. Biomol. Chem., 2014)
One of the rising demands in the field of protein chemical synthesis is the development of facile strategies that yield the protein in workable quantities and homogeneity, with fewer handling steps. , One of the rising demands in the field of protein chemical synthesis is the development of facile strategies that yield the protein in workable quantities and homogeneity, with fewer handling steps. Although the native chemical ligation of peptide hydrazides has recently been shown to be useful for the chemical synthesis of proteins carrying acid-sensitive modification groups, previous hydrazide-based protein synthesis studies have used sequential ligation strategies. Here, we report a practical method for a “one-pot” native chemical ligation of peptide hydrazides that would circumvent the need for the isolation of the intermediate products. This method employed a fast and selective arylboronate oxidation reaction mediated by H 2 O 2 , which draws attention to the potential applications of the thus far under-exploited boron-based functionalities in protein chemical synthesis. To demonstrate the practicality and efficiency of the new one-pot method, we report its application to a scalable total synthesis of modified histones (with five analogues of H3 and H4 as examples) on a multi-milligram scale, with good homogeneity.

One-pot native chemical ligation of peptide hydrazides enables total synthesis of modified histones - Li, Jiabin; Li, Yuanyuan; He, Qiaoqiao; Li, Yiming; Li, Haitao; Liu, Lei (Org. Biomol. Chem., 2014)
One of the rising demands in the field of protein chemical synthesis is the development of facile strategies that yield the protein in workable quantities and homogeneity, with fewer handling steps. , One of the rising demands in the field of protein chemical synthesis is the development of facile strategies that yield the protein in workable quantities and homogeneity, with fewer handling steps. Although the native chemical ligation of peptide hydrazides has recently been shown to be useful for the chemical synthesis of proteins carrying acid-sensitive modification groups, previous hydrazide-based protein synthesis studies have used sequential ligation strategies. Here, we report a practical method for a “one-pot” native chemical ligation of peptide hydrazides that would circumvent the need for the isolation of the intermediate products. This method employed a fast and selective arylboronate oxidation reaction mediated by H 2 O 2 , which draws attention to the potential applications of the thus far under-exploited boron-based functionalities in protein chemical synthesis. To demonstrate the practicality and efficiency of the new one-pot method, we report its application to a scalable total synthesis of modified histones (with five analogues of H3 and H4 as examples) on a multi-milligram scale, with good homogeneity.

Traceless Semisynthesis of a Set of Histone 3 Species Bearing Specific Lysine Methylation Marks - Chen, Zhonglei; Grzybowski, Adrian T.; Ruthenburg, Alexander J. (ChemBioChem, 2014)

Traceless Semisynthesis of a Set of Histone 3 Species Bearing Specific Lysine Methylation Marks - Chen, Zhonglei; Grzybowski, Adrian T.; Ruthenburg, Alexander J. (ChemBioChem, 2014)

Preparation of Fully Synthetic Histone H3 Reveals That Acetyl-Lysine 56 Facilitates Protein Binding Within Nucleosomes - Shimko, John C.; North, Justin A.; Bruns, Aaron N.; Poirier, Michael G.; Ottesen, Jennifer J. (Journal of Molecular Biology, 2011)

Preparation of Fully Synthetic Histone H3 Reveals That Acetyl-Lysine 56 Facilitates Protein Binding Within Nucleosomes - Shimko, John C.; North, Justin A.; Bruns, Aaron N.; Poirier, Michael G.; Ottesen, Jennifer J. (Journal of Molecular Biology, 2011)

5-Methylisoxazole-3-carboxamide-Directed Palladium-Catalyzed γ-C(sp 3 )–H Acetoxylation and Application to the Synthesis of γ-Mercapto Amino Acids for Native Chemical Ligation - Pasunooti, Kalyan Kumar; Yang, Renliang; Banerjee, Biplab; Yap, Terence; Liu, Chuan-Fa (Organic Letters, 2016)

5-Methylisoxazole-3-carboxamide-Directed Palladium-Catalyzed γ-C(sp 3 )–H Acetoxylation and Application to the Synthesis of γ-Mercapto Amino Acids for Native Chemical Ligation - Pasunooti, Kalyan Kumar; Yang, Renliang; Banerjee, Biplab; Yap, Terence; Liu, Chuan-Fa (Organic Letters, 2016)

Total chemical synthesis of methylated analogues of histone 3 revealed KDM4D as a potential regulator of H3K79me3 - Jbara, Muhammad; Guttmann-Raviv, Noga; Maity, Suman Kumar; Ayoub, Nabieh; Brik, Ashraf (Bioorganic & Medicinal Chemistry, 2017)

Total chemical synthesis of methylated analogues of histone 3 revealed KDM4D as a potential regulator of H3K79me3 - Jbara, Muhammad; Guttmann-Raviv, Noga; Maity, Suman Kumar; Ayoub, Nabieh; Brik, Ashraf (Bioorganic & Medicinal Chemistry, 2017)

Examining Several Strategies for the Chemical Synthesis of Phosphorylated Histone H3 Reveals the Effectiveness of the Convergent Approach: Examining Several Strategies for the Chemical Synthesis of Phosphorylated Histone H3 Reveals the Effectiveness of the Convergent Approach - Jbara, Muhammad; Maity, Suman Kumar; Brik, Ashraf (European Journal of Organic Chemistry, 2020)

Examining Several Strategies for the Chemical Synthesis of Phosphorylated Histone H3 Reveals the Effectiveness of the Convergent Approach: Examining Several Strategies for the Chemical Synthesis of Phosphorylated Histone H3 Reveals the Effectiveness of the Convergent Approach - Jbara, Muhammad; Maity, Suman Kumar; Brik, Ashraf (European Journal of Organic Chemistry, 2020)

Total chemical synthesis of bivalently modified H3 by improved three-segment native chemical ligation - Zheng, Yong; Wu, Fangming; Ling, Shenglong; Li, Jia-Bin; Tian, Changlin (Chinese Chemical Letters, 2020)

Total chemical synthesis of bivalently modified H3 by improved three-segment native chemical ligation - Zheng, Yong; Wu, Fangming; Ling, Shenglong; Li, Jia-Bin; Tian, Changlin (Chinese Chemical Letters, 2020)

Chemical Synthesis of Cys-Containing Protein via Chemoselective Deprotection with Different Palladium Complexes - Kamo, Naoki; Hayashi, Gosuke; Okamoto, Akimitsu (Organic Letters, 2019)

Chemical Synthesis of Cys-Containing Protein via Chemoselective Deprotection with Different Palladium Complexes - Kamo, Naoki; Hayashi, Gosuke; Okamoto, Akimitsu (Organic Letters, 2019)

A Modular Ligation Strategy for Asymmetric Bivalent Nucleosomes Trimethylated at K36 and K27 - Guidotti, Nora; Lechner, Carolin C.; Bachmann, Andreas L.; Fierz, Beat (ChemBioChem, 2019)

A Modular Ligation Strategy for Asymmetric Bivalent Nucleosomes Trimethylated at K36 and K27 - Guidotti, Nora; Lechner, Carolin C.; Bachmann, Andreas L.; Fierz, Beat (ChemBioChem, 2019)

Highly efficient solid phase synthesis of large polypeptides by iterative ligations of bis(2-sulfanylethyl)amido (SEA) peptide segments - Raibaut, Laurent; Adihou, Hélène; Desmet, Rémi; Delmas, Agnès F.; Aucagne, Vincent; Melnyk, Oleg (Chemical Science, 2013)

Highly efficient solid phase synthesis of large polypeptides by iterative ligations of bis(2-sulfanylethyl)amido (SEA) peptide segments - Raibaut, Laurent; Adihou, Hélène; Desmet, Rémi; Delmas, Agnès F.; Aucagne, Vincent; Melnyk, Oleg (Chemical Science, 2013)

Observing protein interaction dynamics to chemically defined chromatin fibers by colocalization single-molecule fluorescence microscopy - Mivelaz, Maxime; Fierz, Beat (Methods, 2020)

Crystal structure of the nucleosome containing histone H3 with crotonylated lysine 122 - Suzuki, Yuya; Horikoshi, Naoki; Kato, Daiki; Kurumizaka, Hitoshi (Biochemical and Biophysical Research Communications, 2016)

Observing protein interaction dynamics to chemically defined chromatin fibers by colocalization single-molecule fluorescence microscopy - Mivelaz, Maxime; Fierz, Beat (Methods, 2020)

Chemical Synthesis of O-Glycosylated Human Interleukin-2 by the Reverse Polarity Protection Strategy - Asahina, Yuya; Komiya, Shinobu; Ohagi, Ami; Fujimoto, Rina; Tamagaki, Hiroko; Nakagawa, Katsuhiro; Sato, Takashi; Akira, Shizuo; Takao, Toshifumi; Ishii, Akira; Nakahara, Yoshiaki; Hojo, Hironobu (Angewandte Chemie International Edition, 2015)

Crystal structure of the nucleosome containing histone H3 with crotonylated lysine 122 - Suzuki, Yuya; Horikoshi, Naoki; Kato, Daiki; Kurumizaka, Hitoshi (Biochemical and Biophysical Research Communications, 2016)

Chemical Synthesis of the Highly Hydrophobic Antiviral Membrane-Associated Protein IFITM3 and Modified Variants - Harmand, Thibault J.; Pattabiraman, Vijaya R.; Bode, Jeffrey W. (Angewandte Chemie International Edition, 2017)

Chemical Synthesis of O-Glycosylated Human Interleukin-2 by the Reverse Polarity Protection Strategy - Asahina, Yuya; Komiya, Shinobu; Ohagi, Ami; Fujimoto, Rina; Tamagaki, Hiroko; Nakagawa, Katsuhiro; Sato, Takashi; Akira, Shizuo; Takao, Toshifumi; Ishii, Akira; Nakahara, Yoshiaki; Hojo, Hironobu (Angewandte Chemie International Edition, 2015)

Chemical Synthesis of the Highly Hydrophobic Antiviral Membrane-Associated Protein IFITM3 and Modified Variants - Harmand, Thibault J.; Pattabiraman, Vijaya R.; Bode, Jeffrey W. (Angewandte Chemie International Edition, 2017)

Chemical Synthesis of Interleukin‐2 and Disulfide Stabilizing Analogues - Murar, Claudia E.; Ninomiya, Mamiko; Shimura, Satomi; Karakus, Ufuk; Boyman, Onur; Bode, Jeffrey W. (Angewandte Chemie International Edition, 2020)

The New Salicylaldehyde S , S -Propanedithioacetal Ester Enables N-to-C Sequential Native Chemical Ligation and Ser/Thr Ligation for Chemical Protein Synthesis - Huang, Dong-Liang; Li, Ying; Liang, Jun; Yu, Lu; Xue, Min; Cao, Xiu-Xiu; Xiao, Bin; Tian, Chang-Lin; Liu, Lei; Zheng, Ji-Shen (Journal of the American Chemical Society, 2020)

Chemical Synthesis of Interleukin‐2 and Disulfide Stabilizing Analogues - Murar, Claudia E.; Ninomiya, Mamiko; Shimura, Satomi; Karakus, Ufuk; Boyman, Onur; Bode, Jeffrey W. (Angewandte Chemie International Edition, 2020)

Total chemical synthesis of N-myristoylated HIV-1 matrix protein p17: Structural and mechanistic implications of p17 myristoylation - Wu, Z.; Alexandratos, J.; Ericksen, B.; Lubkowski, J.; Gallo, R. C.; Lu, W. (Proceedings of the National Academy of Sciences, 2004)

The New Salicylaldehyde S , S -Propanedithioacetal Ester Enables N-to-C Sequential Native Chemical Ligation and Ser/Thr Ligation for Chemical Protein Synthesis - Huang, Dong-Liang; Li, Ying; Liang, Jun; Yu, Lu; Xue, Min; Cao, Xiu-Xiu; Xiao, Bin; Tian, Chang-Lin; Liu, Lei; Zheng, Ji-Shen (Journal of the American Chemical Society, 2020)

Total chemical synthesis of N-myristoylated HIV-1 matrix protein p17: Structural and mechanistic implications of p17 myristoylation - Wu, Z.; Alexandratos, J.; Ericksen, B.; Lubkowski, J.; Gallo, R. C.; Lu, W. (Proceedings of the National Academy of Sciences, 2004)

Chemical synthesis of the ATAD2 bromodomain - Creech, G. S.; Paresi, C.; Li, Y.-M.; Danishefsky, S. J. (Proceedings of the National Academy of Sciences, 2014)

Chemical synthesis of the ATAD2 bromodomain - Creech, G. S.; Paresi, C.; Li, Y.-M.; Danishefsky, S. J. (Proceedings of the National Academy of Sciences, 2014)

Convergent chemical synthesis and high-resolution x-ray structure of human lysozyme - Durek, T.; Torbeev, V. Yu.; Kent, S. B. H. (Proceedings of the National Academy of Sciences, 2007)

Rapid Total Synthesis of DARPin pE59 and Barnase - Mong, Surin K.; Vinogradov, Alexander A.; Simon, Mark D.; Pentelute, Bradley L. (ChemBioChem, 2014)

Convergent chemical synthesis and high-resolution x-ray structure of human lysozyme - Durek, T.; Torbeev, V. Yu.; Kent, S. B. H. (Proceedings of the National Academy of Sciences, 2007)

Chemical synthesis of histone H2A with methylation at Gln104 - He, Qiaoqiao; Li, Jiabin; Qi, Yunkun; Wang, Zhipeng; Huang, Yong; Liu, Lei (Science China Chemistry, 2017)

Chemical Synthesis of Phosphorylated Histone H2A at Tyr57 Reveals Insight into the Inhibition Mode of the SAGA Deubiquitinating Module - Jbara, Muhammad; Maity, Suman Kumar; Morgan, Michael; Wolberger, Cynthia; Brik, Ashraf (Angewandte Chemie International Edition, 2016)

Site‐Specific Hyperphosphorylation Inhibits, Rather than Promotes, Tau Fibrillization, Seeding Capacity, and Its Microtubule Binding - Haj‐Yahya, Mahmood; Gopinath, Pushparathinam; Rajasekhar, Kolla; Mirbaha, Hilda; Diamond, Marc I.; Lashuel, Hilal A. (Angewandte Chemie International Edition, 2020)

Rapid Total Synthesis of DARPin pE59 and Barnase - Mong, Surin K.; Vinogradov, Alexander A.; Simon, Mark D.; Pentelute, Bradley L. (ChemBioChem, 2014)

Efficient Chemical Protein Synthesis using Fmoc‐Masked N‐Terminal Cysteine in Peptide Thioester Segments - Kar, Abhisek; Mannuthodikayil, Jamsad; Singh, Sameer; Biswas, Anamika; Dubey, Puneet; Das, Amit; Mandal, Kalyaneswar (Angewandte Chemie International Edition, 2020)

In vitro and in cell analysis of chemically synthesized histone H2A with multiple modifications - Hayashi, Gosuke; Sueoka, Takuma; Okamoto, Akimitsu (Chemical Communications, 2016)
The chemical synthetic route to histone H2A is described. An H2A–H2B dimer, histone octamer, and nucleosome were reconstituted with the synthetic H2A. , The chemical synthetic route to histone H2A is described. An H2A–H2B dimer, histone octamer, and nucleosome were reconstituted with the synthetic H2A. Fluorescein-labeled H2A and multiply modified H2A, which has three different posttranslational modifications, were also synthesized, and applied to live-cell imaging and in vitro nucleosome stability assays, respectively.

Chemical synthesis of histone H2A with methylation at Gln104 - He, Qiaoqiao; Li, Jiabin; Qi, Yunkun; Wang, Zhipeng; Huang, Yong; Liu, Lei (Science China Chemistry, 2017)

P−B Desulfurization: An Enabling Method for Protein Chemical Synthesis and Site-Specific Deuteration - Jin, Kang; Li, Tianlu; Chow, Hoi Yee; Liu, Han; Li, Xuechen (Angewandte Chemie International Edition, 2017)

Chemical Synthesis of Phosphorylated Histone H2A at Tyr57 Reveals Insight into the Inhibition Mode of the SAGA Deubiquitinating Module - Jbara, Muhammad; Maity, Suman Kumar; Morgan, Michael; Wolberger, Cynthia; Brik, Ashraf (Angewandte Chemie International Edition, 2016)

Regulation of the Stability of the Histone H2A–H2B Dimer by H2A Tyr57 Phosphorylation - Sueoka, Takuma; Hayashi, Gosuke; Okamoto, Akimitsu (Biochemistry, 2017)

Site‐Specific Hyperphosphorylation Inhibits, Rather than Promotes, Tau Fibrillization, Seeding Capacity, and Its Microtubule Binding - Haj‐Yahya, Mahmood; Gopinath, Pushparathinam; Rajasekhar, Kolla; Mirbaha, Hilda; Diamond, Marc I.; Lashuel, Hilal A. (Angewandte Chemie International Edition, 2020)

Glycopeptide Synthesis Based on a TFA-Labile Protection Strategy and One-Pot Four-Segment Ligation for the Synthesis of O-Glycosylated Histone H2A: Glycopeptide Synthesis Based on a TFA-Labile Protection Strategy and One-Pot Four-Segment Ligation for the Synthesis of O-Glycosylated Histone H2A - Asahina, Yuya; Kawakami, Toru; Hojo, Hironobu (European Journal of Organic Chemistry, 2019)

Chemical Synthesis of a Synthetic Analogue of the Sialic Acid-Binding Lectin Siglec-7 - Izumi, Masayuki; Otsuki, Akihisa; Nishihara, Mika; Okamoto, Ryo; Kajihara, Yasuhiro (ChemBioChem, 2014)

Efficient Chemical Protein Synthesis using Fmoc‐Masked N‐Terminal Cysteine in Peptide Thioester Segments - Kar, Abhisek; Mannuthodikayil, Jamsad; Singh, Sameer; Biswas, Anamika; Dubey, Puneet; Das, Amit; Mandal, Kalyaneswar (Angewandte Chemie International Edition, 2020)

Cysteinylprolyl imide (CPI) peptide: a highly reactive and easily accessible crypto-thioester for chemical protein synthesis - Yanase, Masafumi; Nakatsu, Koki; Cardos, Charlane Joy; Konda, Yoshiki; Hayashi, Gosuke; Okamoto, Akimitsu (Chemical Science, 2019)
A new crypto-thioester, cysteinylprolyl imide (CPI) peptide, offers a practical synthetic pathway and reliable reaction rate to be successfully applied to chemical protein synthesis. , Native chemical ligation (NCL) between the C-terminal peptide thioester and the N-terminal cysteinyl-peptide revolutionized the field of chemical protein synthesis. The difficulty of direct synthesis of the peptide thioester in the Fmoc method has prompted the development of crypto-thioesters that can be efficiently converted into thioesters. Cysteinylprolyl ester (CPE), which is an N – S acyl shift-driven crypto-thioester that relies on an intramolecular O – N acyl shift to displace the amide-thioester equilibrium, enabled trans -thioesterification and subsequent NCL in one pot. However, the utility of CPE is limited because of the moderate thioesterification rates and the synthetic complexity introduced by the ester group. Herein, we develop a new crypto-thioester, cysteinylprolyl imide (CPI), which replaces the alcohol leaving group of CPE with other leaving groups such as benzimidazolidinone, oxazolidinone, and pyrrolidinone. CPI peptides were efficiently synthesized by using standard Fmoc solid-phase peptide synthesis (SPPS) and subsequent on-resin imide formation. Screening of the several imide structures indicated that methyloxazolidinone- t -leucine (MeOxd-Tle) showed faster conversion into thioester and higher stability against hydrolysis under NCL conditions. Finally, by using CPMeOxd-Tle peptides, we demonstrated the chemical synthesis of affibody via N-to-C sequential, three-segment ligation and histone H2A.Z via convergent four-segment ligation. This facile and straightforward method is expected to be broadly applicable to chemical protein synthesis.

In vitro and in cell analysis of chemically synthesized histone H2A with multiple modifications - Hayashi, Gosuke; Sueoka, Takuma; Okamoto, Akimitsu (Chemical Communications, 2016)
The chemical synthetic route to histone H2A is described. An H2A–H2B dimer, histone octamer, and nucleosome were reconstituted with the synthetic H2A. , The chemical synthetic route to histone H2A is described. An H2A–H2B dimer, histone octamer, and nucleosome were reconstituted with the synthetic H2A. Fluorescein-labeled H2A and multiply modified H2A, which has three different posttranslational modifications, were also synthesized, and applied to live-cell imaging and in vitro nucleosome stability assays, respectively.

P−B Desulfurization: An Enabling Method for Protein Chemical Synthesis and Site-Specific Deuteration - Jin, Kang; Li, Tianlu; Chow, Hoi Yee; Liu, Han; Li, Xuechen (Angewandte Chemie International Edition, 2017)

Regulation of the Stability of the Histone H2A–H2B Dimer by H2A Tyr57 Phosphorylation - Sueoka, Takuma; Hayashi, Gosuke; Okamoto, Akimitsu (Biochemistry, 2017)

Chemical Synthesis of the Homogeneous Granulocyte-Macrophage Colony-Stimulating Factor Through Se-Auxiliary-Mediated Ligation - Lu, Dan; Yin, Hongli; Wang, Siyao; Tang, Feng; Huang, Wei; Wang, Ping (The Journal of Organic Chemistry, 2020)

Glycopeptide Synthesis Based on a TFA-Labile Protection Strategy and One-Pot Four-Segment Ligation for the Synthesis of O-Glycosylated Histone H2A: Glycopeptide Synthesis Based on a TFA-Labile Protection Strategy and One-Pot Four-Segment Ligation for the Synthesis of O-Glycosylated Histone H2A - Asahina, Yuya; Kawakami, Toru; Hojo, Hironobu (European Journal of Organic Chemistry, 2019)

Total chemical synthesis of human T-cell leukemia virus type 1 protease via native chemical ligation - Li, Changqing; Li, Xiangqun; Lu, Wuyuan (Biopolymers, 2010)

Chemical Synthesis of a Synthetic Analogue of the Sialic Acid-Binding Lectin Siglec-7 - Izumi, Masayuki; Otsuki, Akihisa; Nishihara, Mika; Okamoto, Ryo; Kajihara, Yasuhiro (ChemBioChem, 2014)

Cysteinylprolyl imide (CPI) peptide: a highly reactive and easily accessible crypto-thioester for chemical protein synthesis - Yanase, Masafumi; Nakatsu, Koki; Cardos, Charlane Joy; Konda, Yoshiki; Hayashi, Gosuke; Okamoto, Akimitsu (Chemical Science, 2019)
A new crypto-thioester, cysteinylprolyl imide (CPI) peptide, offers a practical synthetic pathway and reliable reaction rate to be successfully applied to chemical protein synthesis. , Native chemical ligation (NCL) between the C-terminal peptide thioester and the N-terminal cysteinyl-peptide revolutionized the field of chemical protein synthesis. The difficulty of direct synthesis of the peptide thioester in the Fmoc method has prompted the development of crypto-thioesters that can be efficiently converted into thioesters. Cysteinylprolyl ester (CPE), which is an N – S acyl shift-driven crypto-thioester that relies on an intramolecular O – N acyl shift to displace the amide-thioester equilibrium, enabled trans -thioesterification and subsequent NCL in one pot. However, the utility of CPE is limited because of the moderate thioesterification rates and the synthetic complexity introduced by the ester group. Herein, we develop a new crypto-thioester, cysteinylprolyl imide (CPI), which replaces the alcohol leaving group of CPE with other leaving groups such as benzimidazolidinone, oxazolidinone, and pyrrolidinone. CPI peptides were efficiently synthesized by using standard Fmoc solid-phase peptide synthesis (SPPS) and subsequent on-resin imide formation. Screening of the several imide structures indicated that methyloxazolidinone- t -leucine (MeOxd-Tle) showed faster conversion into thioester and higher stability against hydrolysis under NCL conditions. Finally, by using CPMeOxd-Tle peptides, we demonstrated the chemical synthesis of affibody via N-to-C sequential, three-segment ligation and histone H2A.Z via convergent four-segment ligation. This facile and straightforward method is expected to be broadly applicable to chemical protein synthesis.

VHH characterization. Comparison of recombinant with chemically synthesized anti‐HER2 VHH - Hartmann, Lucie; Botzanowski, Thomas; Galibert, Mathieu; Jullian, Magali; Chabrol, Eric; Zeder‐Lutz, Gabrielle; Kugler, Valérie; Stojko, Johann; Strub, Jean‐Marc; Ferry, Gilles; Frankiewicz, Lukasz; Puget, Karine; et al. (Protein Science, 2019)

Chemical Synthesis of the Homogeneous Granulocyte-Macrophage Colony-Stimulating Factor Through Se-Auxiliary-Mediated Ligation - Lu, Dan; Yin, Hongli; Wang, Siyao; Tang, Feng; Huang, Wei; Wang, Ping (The Journal of Organic Chemistry, 2020)

Protein Chemical Synthesis by Ligation of Peptide Hydrazides - Fang, Ge-Min; Li, Yi-Ming; Shen, Fei; Huang, Yi-Chao; Li, Jia-Bin; Lin, Yun; Cui, Hong-Kui; Liu, Lei (Angewandte Chemie International Edition, 2011)

Total chemical synthesis of human T-cell leukemia virus type 1 protease via native chemical ligation - Li, Changqing; Li, Xiangqun; Lu, Wuyuan (Biopolymers, 2010)

Semisynthesis and Folding of the Potassium Channel KcsA - Valiyaveetil, Francis I.; MacKinnon, Roderick; Muir, Tom W. (Journal of the American Chemical Society, 2002)

Peptide o -Aminoanilides as Crypto-Thioesters for Protein Chemical Synthesis - Wang, Jia-Xing; Fang, Ge-Min; He, Yao; Qu, Da-Liang; Yu, Min; Hong, Zhang-Yong; Liu, Lei (Angewandte Chemie International Edition, 2015)

VHH characterization. Comparison of recombinant with chemically synthesized anti‐HER2 VHH - Hartmann, Lucie; Botzanowski, Thomas; Galibert, Mathieu; Jullian, Magali; Chabrol, Eric; Zeder‐Lutz, Gabrielle; Kugler, Valérie; Stojko, Johann; Strub, Jean‐Marc; Ferry, Gilles; Frankiewicz, Lukasz; Puget, Karine; et al. (Protein Science, 2019)

Total chemical synthesis of fully functional Photoactive Yellow Protein - Gordon, Wendy R.; Bang, Duhee; Hoff, Wouter D.; Kent, Stephen B.H. (Bioorganic & Medicinal Chemistry, 2013)

Protein Chemical Synthesis by Ligation of Peptide Hydrazides - Fang, Ge-Min; Li, Yi-Ming; Shen, Fei; Huang, Yi-Chao; Li, Jia-Bin; Lin, Yun; Cui, Hong-Kui; Liu, Lei (Angewandte Chemie International Edition, 2011)

A Semisynthetic Strategy to Generate Phosphorylated and Acetylated Histone H2B - Chiang, Kyle P.; Jensen, Mette S.; McGinty, Robert K.; Muir, Tom W. (ChemBioChem, 2009)

Semisynthesis and Folding of the Potassium Channel KcsA - Valiyaveetil, Francis I.; MacKinnon, Roderick; Muir, Tom W. (Journal of the American Chemical Society, 2002)

Peptide o -Aminoanilides as Crypto-Thioesters for Protein Chemical Synthesis - Wang, Jia-Xing; Fang, Ge-Min; He, Yao; Qu, Da-Liang; Yu, Min; Hong, Zhang-Yong; Liu, Lei (Angewandte Chemie International Edition, 2015)

Chemical Synthesis of Proteins with Non-Strategically Placed Cysteines Using Selenazolidine and Selective Deselenization - Reddy, Post Sai; Dery, Shahar; Metanis, Norman (Angewandte Chemie International Edition, 2016)

Total chemical synthesis of fully functional Photoactive Yellow Protein - Gordon, Wendy R.; Bang, Duhee; Hoff, Wouter D.; Kent, Stephen B.H. (Bioorganic & Medicinal Chemistry, 2013)

Protein Assembly by Orthogonal Chemical Ligation Methods - Nilsson, Bradley L.; Hondal, Robert J.; Soellner, Matthew B.; Raines, Ronald T. (Journal of the American Chemical Society, 2003)

A Semisynthetic Strategy to Generate Phosphorylated and Acetylated Histone H2B - Chiang, Kyle P.; Jensen, Mette S.; McGinty, Robert K.; Muir, Tom W. (ChemBioChem, 2009)

Conformationally Assisted Protein Ligation Using C-Terminal Thioester Peptides - Beligere, Gangamani S.; Dawson, Philip E. (Journal of the American Chemical Society, 1999)

Protein synthesis by native chemical ligation: Expanded scope by using straightforward methodology - Hackeng, T. M.; Griffin, J. H.; Dawson, P. E. (Proceedings of the National Academy of Sciences, 1999)

Chemical Synthesis of Proteins with Non-Strategically Placed Cysteines Using Selenazolidine and Selective Deselenization - Reddy, Post Sai; Dery, Shahar; Metanis, Norman (Angewandte Chemie International Edition, 2016)

Protein Assembly by Orthogonal Chemical Ligation Methods - Nilsson, Bradley L.; Hondal, Robert J.; Soellner, Matthew B.; Raines, Ronald T. (Journal of the American Chemical Society, 2003)

Conformationally Assisted Protein Ligation Using C-Terminal Thioester Peptides - Beligere, Gangamani S.; Dawson, Philip E. (Journal of the American Chemical Society, 1999)

Total synthesis by modern chemical ligation methods and high resolution (1.1 Å) X-ray structure of ribonuclease A - Boerema, David J.; Tereshko, Valentina A.; Kent, Stephen B. H. (Biopolymers, 2008)

Protein synthesis by native chemical ligation: Expanded scope by using straightforward methodology - Hackeng, T. M.; Griffin, J. H.; Dawson, P. E. (Proceedings of the National Academy of Sciences, 1999)

Selenocysteine in Native Chemical Ligation and Expressed Protein Ligation - Hondal, Robert J.; Nilsson, Bradley L.; Raines, Ronald T. (Journal of the American Chemical Society, 2001)

Semisynthesis of a Homogeneous Glycoprotein Enzyme: Ribonuclease C: Part 2 - Piontek, Christian; Varón Silva, Daniel; Heinlein, Christian; Pöhner, Claudia; Mezzato, Stefano; Ring, Petra; Martin, Andreas; Schmid, Franz Xaver; Unverzagt, Carlo (Angewandte Chemie International Edition, 2009)

Total synthesis by modern chemical ligation methods and high resolution (1.1 Å) X-ray structure of ribonuclease A - Boerema, David J.; Tereshko, Valentina A.; Kent, Stephen B. H. (Biopolymers, 2008)

Chemical synthesis and enzymatic properties of RNase A analogues designed to enhance second-step catalytic activity - Boerema, David J.; Tereshko, Valentina A.; Zhang, Junliang; Kent, Stephen B. H. (Organic & Biomolecular Chemistry, 2016)
Adenine covalently attached to the RNase A enzyme molecule decreased the rate of transphosphorylation and increased the rate of hydrolysis. , In this paper, we have used total chemical synthesis of RNase A analogues in order to probe the molecular basis of enzyme catalysis. Our goal was to obligately fill the adenine-binding pocket on the enzyme molecule, and to thus pre-orient the imidazole side chain of His 119 in its catalytically productive orientation. Two designed analogues of the RNase A protein molecule that contained an adenine moiety covalently bound to distinct amino acid side chains adjacent to the adenine binding pocket were prepared. A crystal structure of one analogue was determined at 2.3 Å resolution. Kinetic data for RNA transphosporylation and 2′,3′ cyclic mononucleotide hydrolysis were acquired for the adenine-containing RNase A analogue proteins. As anticipated, the presence of a covalently attached adenine on the enzyme molecule decreased the rate of transphosphorylation and increased the rate of hydrolysis, although the magnitude of the effects was small. This work illustrates the use of total protein synthesis to investigate the chemistry of enzyme catalysis in ways not possible through traditional biochemistry or molecular biology.

Selenocysteine in Native Chemical Ligation and Expressed Protein Ligation - Hondal, Robert J.; Nilsson, Bradley L.; Raines, Ronald T. (Journal of the American Chemical Society, 2001)

Correct Disulfide Pairing Is Required for the Biological Activity of Crustacean Androgenic Gland Hormone (AGH): Synthetic Studies of AGH - Katayama, Hidekazu; Hojo, Hironobu; Ohira, Tsuyoshi; Ishii, Akira; Nozaki, Takamichi; Goto, Kiyomi; Nakahara, Yuko; Takahashi, Tetsuo; Hasegawa, Yuriko; Nagasawa, Hiromichi; Nakahara, Yoshiaki (Biochemistry, 2010)

Semisynthesis of a Homogeneous Glycoprotein Enzyme: Ribonuclease C: Part 2 - Piontek, Christian; Varón Silva, Daniel; Heinlein, Christian; Pöhner, Claudia; Mezzato, Stefano; Ring, Petra; Martin, Andreas; Schmid, Franz Xaver; Unverzagt, Carlo (Angewandte Chemie International Edition, 2009)

Peptidyl N , N -Bis(2-mercaptoethyl)-amides as Thioester Precursors for Native Chemical Ligation † - Hou, Wen; Zhang, Xiaohong; Li, Fupeng; Liu, Chuan-Fa (Organic Letters, 2011)

Chemical synthesis and enzymatic properties of RNase A analogues designed to enhance second-step catalytic activity - Boerema, David J.; Tereshko, Valentina A.; Zhang, Junliang; Kent, Stephen B. H. (Organic & Biomolecular Chemistry, 2016)
Adenine covalently attached to the RNase A enzyme molecule decreased the rate of transphosphorylation and increased the rate of hydrolysis. , In this paper, we have used total chemical synthesis of RNase A analogues in order to probe the molecular basis of enzyme catalysis. Our goal was to obligately fill the adenine-binding pocket on the enzyme molecule, and to thus pre-orient the imidazole side chain of His 119 in its catalytically productive orientation. Two designed analogues of the RNase A protein molecule that contained an adenine moiety covalently bound to distinct amino acid side chains adjacent to the adenine binding pocket were prepared. A crystal structure of one analogue was determined at 2.3 Å resolution. Kinetic data for RNA transphosporylation and 2′,3′ cyclic mononucleotide hydrolysis were acquired for the adenine-containing RNase A analogue proteins. As anticipated, the presence of a covalently attached adenine on the enzyme molecule decreased the rate of transphosphorylation and increased the rate of hydrolysis, although the magnitude of the effects was small. This work illustrates the use of total protein synthesis to investigate the chemistry of enzyme catalysis in ways not possible through traditional biochemistry or molecular biology.

Total Chemical Synthesis of an Integral Membrane Enzyme: Diacylglycerol Kinase from Escherichia coli - Lahiri, Sunanda; Brehs, Manuel; Olschewski, Diana; Becker, Christian F. W. (Angewandte Chemie International Edition, 2011)

Correct Disulfide Pairing Is Required for the Biological Activity of Crustacean Androgenic Gland Hormone (AGH): Synthetic Studies of AGH - Katayama, Hidekazu; Hojo, Hironobu; Ohira, Tsuyoshi; Ishii, Akira; Nozaki, Takamichi; Goto, Kiyomi; Nakahara, Yuko; Takahashi, Tetsuo; Hasegawa, Yuriko; Nagasawa, Hiromichi; Nakahara, Yoshiaki (Biochemistry, 2010)

Expedient synthesis of coumarin-coupled triazoles via ‘click chemistry’ leading to the formation of coumarin–triazole–sugar hybrids - Kumar, K. Karthik; Kumar, R. Mahesh; Subramanian, V.; Das, T. Mohan (Carbohydrate Research, 2010)

Peptidyl N , N -Bis(2-mercaptoethyl)-amides as Thioester Precursors for Native Chemical Ligation † - Hou, Wen; Zhang, Xiaohong; Li, Fupeng; Liu, Chuan-Fa (Organic Letters, 2011)

A Central Cysteine Residue Is Essential for the Thermal Stability and Function of SUMO-1 Protein and SUMO-1 Peptide–Protein Conjugates - Drobecq, Hervé; Boll, Emmanuelle; Sénéchal, Magalie; Desmet, Rémi; Saliou, Jean-Michel; Lacapère, Jean-Jacques; Mougel, Alexandra; Vicogne, Jérôme; Melnyk, Oleg (Bioconjugate Chemistry, 2016)

Total Chemical Synthesis of an Integral Membrane Enzyme: Diacylglycerol Kinase from Escherichia coli - Lahiri, Sunanda; Brehs, Manuel; Olschewski, Diana; Becker, Christian F. W. (Angewandte Chemie International Edition, 2011)

Expedient synthesis of coumarin-coupled triazoles via ‘click chemistry’ leading to the formation of coumarin–triazole–sugar hybrids - Kumar, K. Karthik; Kumar, R. Mahesh; Subramanian, V.; Das, T. Mohan (Carbohydrate Research, 2010)

A Central Cysteine Residue Is Essential for the Thermal Stability and Function of SUMO-1 Protein and SUMO-1 Peptide–Protein Conjugates - Drobecq, Hervé; Boll, Emmanuelle; Sénéchal, Magalie; Desmet, Rémi; Saliou, Jean-Michel; Lacapère, Jean-Jacques; Mougel, Alexandra; Vicogne, Jérôme; Melnyk, Oleg (Bioconjugate Chemistry, 2016)

Chemical synthesis of γ-secretase activating protein using pseudoglutamines as ligation sites: Synthesis of γ-Secretase Activating Protein - Harris, Paul W. R.; Squire, Chris; Young, Paul G.; Brimble, Margaret A. (Biopolymers, 2015)

A Novel d ‐Peptide Identified by Mirror‐Image Phage Display Blocks TIGIT/PVR for Cancer Immunotherapy - Zhou, Xiuman; Zuo, Chao; Li, Wanqiong; Shi, Weiwei; Zhou, Xiaowen; Wang, Hongfei; Chen, Shaomeng; Du, Jiangfeng; Chen, Guanyu; Zhai, Wenjie; Zhao, Wenshan; Wu, Yahong; et al. (Angewandte Chemie International Edition, 2020)

Chemical Protein Synthesis by Solid Phase Ligation of Unprotected Peptide Segments - Canne, Lynne E.; Botti, Paolo; Simon, Reyna J.; Chen, Yijun; Dennis, Edward A.; Kent, Stephen B. H. (Journal of the American Chemical Society, 1999)

Total synthesis of TRADD death domain with arginine N-GlcNAcylation by hydrazide-based native chemical ligation - Wu, Ye; Li, Yulei; Cong, Wei; Zou, Yan; Li, Xiang; Hu, Honggang (Chinese Chemical Letters, 2020)

Total chemical synthesis of an Orf virus protein, ORFV002, an inhibitor of the master gene regulator NF-κB: Chemical Synthesis of ORFV002 - Son, S. J.; Harris, P. W. R.; Squire, C. J.; Baker, E. N.; Kent, S. B. H.; Brimble, M. A. (Biopolymers, 2014)

Chemical Synthesis and Expression of the HIV-1 Rev Protein - Siman, Peter; Blatt, Ofrah; Moyal, Tal; Danieli, Tsafi; Lebendiker, Mario; Lashuel, Hilal A.; Friedler, Assaf; Brik, Ashraf (ChemBioChem, 2011)

Chemical synthesis of γ-secretase activating protein using pseudoglutamines as ligation sites: Synthesis of γ-Secretase Activating Protein - Harris, Paul W. R.; Squire, Chris; Young, Paul G.; Brimble, Margaret A. (Biopolymers, 2015)

A Novel d ‐Peptide Identified by Mirror‐Image Phage Display Blocks TIGIT/PVR for Cancer Immunotherapy - Zhou, Xiuman; Zuo, Chao; Li, Wanqiong; Shi, Weiwei; Zhou, Xiaowen; Wang, Hongfei; Chen, Shaomeng; Du, Jiangfeng; Chen, Guanyu; Zhai, Wenjie; Zhao, Wenshan; Wu, Yahong; et al. (Angewandte Chemie International Edition, 2020)