Semienzymatic cyclization of disulfide-rich peptides using sortase A

Jia, Xinying, Kwon, Soohyun, Wang, Ching-I Anderson, Huang, Yen-Hua, Chan, Lai Y., Tan, Chia Chia, Rosengren, K. Johan, Mulvenna, Jason P., Schroeder, Christina I. and Craik, David J. (2014) Semienzymatic cyclization of disulfide-rich peptides using sortase A. Journal of Biological Chemistry, Papers in Press 10: 1-25. doi:10.1074/jbc.M113.539262

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Author Jia, Xinying
Kwon, Soohyun
Wang, Ching-I Anderson
Huang, Yen-Hua
Chan, Lai Y.
Tan, Chia Chia
Rosengren, K. Johan
Mulvenna, Jason P.
Schroeder, Christina I.
Craik, David J.
Title Semienzymatic cyclization of disulfide-rich peptides using sortase A
Journal name Journal of Biological Chemistry   Check publisher's open access policy
ISSN 0021-9258
Publication date 2014-01-14
Year available 2014
Sub-type Article (original research)
DOI 10.1074/jbc.M113.539262
Open Access Status File (Author Post-print)
Volume Papers in Press
Issue 10
Start page 1
End page 25
Total pages 25
Place of publication Bethesda, MD, United States
Publisher American Society for Biochemistry and Molecular Biology
Language eng
Formatted abstract
Disulfide-rich cyclic peptides have generated great interest in the development of peptide-based therapeutics due to their exceptional stability towards chemical, enzymatic or thermal attack. In particular, they have been used as scaffolds onto which bioactive epitopes can be grafted to take advantage of the favorable biophysical properties of disulfide-rich cyclic peptides. To date, the most commonly used method for the head-to-tail cyclization of peptides has been native chemical ligation. In recent years, however, enzyme-mediated cyclization has become a promising new technology due to its efficiency, safety and cost-effectiveness. Sortase A (SrtA) is a bacterial enzyme with transpeptidase activity. It recognizes a C-terminal penta-amino acid motif ′LPXTG′ and cleaves the amide bond between Thr and Gly to form a thioacyl-linked intermediate. This intermediate undergoes nucleophilic attack by an N-terminal poly-Gly sequence to form an amide bond between the Thr and N-terminal Gly. Here, we demonstrate that Sortase A can successfully be used to cyclize a variety of small disulfide-rich peptides, including the cyclotide kalata B1, α-conotoxin Vc1.1 and sunflower trypsin inhibitor 1 (SFTI-1). These peptides range in size from 14 to 29 amino acids and respectively contain three, two or one disulfide bond within their head-to-tail cyclic backbones. Our findings provide proof-of-concept for the potential broad applicability of enzymatic cyclization of disulfide-rich peptides with therapeutic potential.

Background: Sortase A (SrtA) is a transpeptidase capable of catalyzing the formation of amide bonds.
Results: SrtA was used to backbone cyclize disulfide-rich peptides including kalata B1, α-conotoxin Vc1.1 and SFTI-1.
Conclusion: SrtA-mediated cyclization is applicable to small disulfide-rich peptides.
Significance: SrtA-mediated cyclization is an alternative to native chemical ligation for the cyclization of small peptides of therapeutic interest.
Keyword Disulfide
Peptide chemical synthesis
Cyclic peptides
Enzymatic cyclization
Kalata B1
Sortase A
Q-Index Code C1
Q-Index Status Confirmed Code
Institutional Status UQ
Additional Notes First Published on January 14, 2014

Document type: Journal Article
Sub-type: Article (original research)
Collections: Official 2015 Collection
School of Biomedical Sciences Publications
Institute for Molecular Bioscience - Publications
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Citation counts: TR Web of Science Citation Count  Cited 30 times in Thomson Reuters Web of Science Article | Citations
Scopus Citation Count Cited 35 times in Scopus Article | Citations
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Created: Thu, 13 Feb 2014, 00:54:24 EST by Susan Allen on behalf of Institute for Molecular Bioscience