Screening for expressed nonribosomal peptide synthetases and polyketide synthases using LC-MS/MS-based proteomics

Yunqiu Chen, Ryan A. McClure, Neil L. Kelleher

Research output: Chapter in Book/Report/Conference proceedingChapter

4 Scopus citations

Abstract

Liquid chromatography–mass spectrometry (LC-MS)-based proteomics is a powerful technique for the profi ling of protein expression in cells in a high-throughput fashion. Herein we report a protocol using LC-MS/MS-based proteomics for the screening of enzymes involved in natural product biosynthesis, such as nonribosomal peptide synthetases (NRPSs) and polyketide synthases (PKSs) from bacterial strains. Taking advantage of the large size of modular NRPSs and PKSs (often >200 kDa), size-based separation (SDS-PAGE) is employed prior to LC-MS/MS analysis. Based upon the protein identifi cations obtained through software search, we can accurately pinpoint the expressed NRPS and/or PKS gene clusters from a given strain and growth condition. The proteomics screening result can be used to guide the discovery of potentially new nonribosomal peptide and polyketide natural products.

Original languageEnglish (US)
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Pages135-147
Number of pages13
DOIs
StatePublished - Feb 1 2016

Publication series

NameMethods in Molecular Biology
Volume1401
ISSN (Print)1064-3745

Keywords

  • Liquid chromatography
  • Mass spectrometry
  • Natural product
  • Nonribosomal peptide
  • Polyketide
  • Proteomics

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

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    Chen, Y., McClure, R. A., & Kelleher, N. L. (2016). Screening for expressed nonribosomal peptide synthetases and polyketide synthases using LC-MS/MS-based proteomics. In Methods in Molecular Biology (pp. 135-147). (Methods in Molecular Biology; Vol. 1401). Humana Press Inc.. https://doi.org/10.1007/978-1-4939-3375-4_9