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

Akello J Agwa, David J Craik, Christina I Schroeder

Department of Life Sciences

University of Queensland

Research output: Chapter or section in a book/report/conference proceeding › Chapter or section

5 Citations (SciVal)

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.

Original language	English
Title of host publication	Enzyme-Mediated Ligation Methods
Editors	M Schmidt, T Nuijens
Place of Publication	New York
Publisher	Humana Press
Chapter	3
Pages	29-41
Number of pages	13
Volume	2012
ISBN (Electronic)	978-1-4939-9546-2
ISBN (Print)	978-1-4939-9545-5
DOIs	https://doi.org/10.1007/978-1-4939-9546-2_3
Publication status	Published - 30 Jun 2019

Publication series

Name	Methods in Molecular Biology
Volume	2012
ISSN (Print)	1064-3745
ISSN (Electronic)	1940-6029

Keywords

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

ASJC Scopus subject areas

Molecular Biology
Genetics

Access to Document

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

Cite this

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title = "Cyclizing Disulfide-Rich Peptides Using Sortase A",

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",

author = "Agwa, {Akello J} and Craik, {David J} and Schroeder, {Christina I}",

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AU - Craik, David J

AU - Schroeder, Christina I

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N2 - 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.

AB - 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.

KW - Cyclization

KW - Cyclotide

KW - Head-to-tail cyclic peptide

KW - Kalata B1

KW - Macrocycle

KW - Peptide ligation

KW - Semienzymatic

KW - Sortase A

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U2 - 10.1007/978-1-4939-9546-2_3

DO - 10.1007/978-1-4939-9546-2_3

M3 - Chapter or section

C2 - 31161502

SN - 978-1-4939-9545-5

VL - 2012

T3 - Methods in Molecular Biology

SP - 29

EP - 41

BT - Enzyme-Mediated Ligation Methods

A2 - Schmidt, M

A2 - Nuijens, T

PB - Humana Press

CY - New York

ER -

Cyclizing Disulfide-Rich Peptides Using Sortase A

Abstract

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Keywords

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