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Issue 44, 2019
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HAMA: a multiplexed LC-MS/MS assay for specificity profiling of adenylate-forming enzymes

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Abstract

Adenylation enzymes selecting substrates for ribosomal and nonribosomal protein and peptide biosynthesis have been popular targets of enzyme engineering. Previous standard assays for adenylation specificity have been cumbersome and failed to reflect the competition conditions inside a cell because they measure substrates one at a time. We have developed an adenylation assay based on hydroxamate quenching and LC-MS/MS detection of hydroxamate products testing dozens of competing amino acid substrates in parallel. Streamlined specificity profiling of adenylation enzymes will facilitate engineering and directed evolution of ribosomal and nonribosomal peptide synthesis.

Graphical abstract: HAMA: a multiplexed LC-MS/MS assay for specificity profiling of adenylate-forming enzymes

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Supplementary files

Article information


Submitted
22 Aug 2019
Accepted
13 Sep 2019
First published
03 Oct 2019

This article is Open Access
All publication charges for this article have been paid for by the Royal Society of Chemistry

Chem. Sci., 2019,10, 10395-10399
Article type
Edge Article

HAMA: a multiplexed LC-MS/MS assay for specificity profiling of adenylate-forming enzymes

A. Stanišić, A. Hüsken and H. Kries, Chem. Sci., 2019, 10, 10395
DOI: 10.1039/C9SC04222A

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