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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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Publication details

The article was received on 22 Aug 2019, accepted on 13 Sep 2019 and first published on 03 Oct 2019


Article type: Edge Article
DOI: 10.1039/C9SC04222A
Chem. Sci., 2019,10, 10395-10399
  • Open access: Creative Commons BY-NC license
    All publication charges for this article have been paid for by the Royal Society of Chemistry

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    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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