Cyclizing Disulfide-Rich Peptides Using Sortase A

Akello J Agwa; David J Craik; Christina I Schroeder

doi:10.1007/978-1-4939-9546-2_3

Cyclizing Disulfide-Rich Peptides Using Sortase A

Methods Mol Biol. 2019:2012:29-41. doi: 10.1007/978-1-4939-9546-2_3.

Authors

Akello J Agwa¹, David J Craik², Christina I Schroeder³

Affiliations

¹ Institute for Molecular Bioscience, The University of Queensland, Brisbane, QLD, Australia.
² Institute for Molecular Bioscience, The University of Queensland, Brisbane, QLD, Australia. d.craik@imb.uq.edu.au.
³ Institute for Molecular Bioscience, The University of Queensland, Brisbane, QLD, Australia. c.schroeder@imb.uq.edu.au.

PMID: 31161502
DOI: 10.1007/978-1-4939-9546-2_3

Abstract

Sortase A (SrtA) is an enzyme obtained from Staphylococcus aureus that catalyzes site-specific transpeptidation of surface proteins to the bacterial cell membrane. SrtA recognizes an LPXTG amino acid motif and cleaves between the Thr and Gly to form a thioester-linked acyl-enzyme intermediate. The intermediate is resolved in the presence of a nucleophilic N-terminal polyglycine resulting in ligation of the acyl donor to the polyglycine acceptor. Here we describe the application of SrtA as a tool for the cyclization of disulfide-rich peptides. Reactions are typically tailored to each disulfide-rich peptide with optimal conditions producing yields of 40-50% cyclized peptide.

Keywords: Cyclization; Cyclotide; Head-to-tail cyclic peptide; Kalata B1; Macrocycle; Peptide ligation; Semienzymatic; Sortase A.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Amino Acid Motifs
Amino Acid Sequence
Aminoacyltransferases / chemistry*
Bacterial Proteins / chemistry*
Cyclization
Cysteine Endopeptidases / chemistry*
Disulfides / chemistry*
Peptides, Cyclic / chemistry*
Peptides, Cyclic / isolation & purification
Substrate Specificity

Substances

Bacterial Proteins
Disulfides
Peptides, Cyclic
Aminoacyltransferases
sortase A
Cysteine Endopeptidases