Cyclizing Disulfide-Rich Peptides Using Sortase A

Akello J Agwa, David J Craik, Christina I Schroeder

Research output: Chapter in Book/Report/Conference proceedingChapter

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 languageEnglish
Title of host publicationEnzyme-Mediated Ligation Methods
EditorsM Schmidt, T Nuijens
Place of PublicationNew York
PublisherSpringer
Chapter3
Pages29-41
Number of pages13
Volume2012
ISBN (Electronic)978-1-4939-9546-2
ISBN (Print)978-1-4939-9545-5
DOIs
Publication statusPublished - 30 Jun 2019

Publication series

NameMethods in molecular biology (Clifton, N.J.)
PublisherHumana Press
ISSN (Print)1064-3745

Keywords

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

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

Cite this

Agwa, A. J., Craik, D. J., & Schroeder, C. I. (2019). Cyclizing Disulfide-Rich Peptides Using Sortase A. In M. Schmidt, & T. Nuijens (Eds.), Enzyme-Mediated Ligation Methods (Vol. 2012, pp. 29-41). (Methods in molecular biology (Clifton, N.J.)). New York: Springer. https://doi.org/10.1007/978-1-4939-9546-2_3

Cyclizing Disulfide-Rich Peptides Using Sortase A. / Agwa, Akello J; Craik, David J; Schroeder, Christina I.

Enzyme-Mediated Ligation Methods. ed. / M Schmidt; T Nuijens. Vol. 2012 New York : Springer, 2019. p. 29-41 (Methods in molecular biology (Clifton, N.J.)).

Research output: Chapter in Book/Report/Conference proceedingChapter

Agwa, AJ, Craik, DJ & Schroeder, CI 2019, Cyclizing Disulfide-Rich Peptides Using Sortase A. in M Schmidt & T Nuijens (eds), Enzyme-Mediated Ligation Methods. vol. 2012, Methods in molecular biology (Clifton, N.J.), Springer, New York, pp. 29-41. https://doi.org/10.1007/978-1-4939-9546-2_3
Agwa AJ, Craik DJ, Schroeder CI. Cyclizing Disulfide-Rich Peptides Using Sortase A. In Schmidt M, Nuijens T, editors, Enzyme-Mediated Ligation Methods. Vol. 2012. New York: Springer. 2019. p. 29-41. (Methods in molecular biology (Clifton, N.J.)). https://doi.org/10.1007/978-1-4939-9546-2_3
Agwa, Akello J ; Craik, David J ; Schroeder, Christina I. / Cyclizing Disulfide-Rich Peptides Using Sortase A. Enzyme-Mediated Ligation Methods. editor / M Schmidt ; T Nuijens. Vol. 2012 New York : Springer, 2019. pp. 29-41 (Methods in molecular biology (Clifton, N.J.)).
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