ArticleMethods in molecular biology (Clifton, N.J.)2023
Thioester Capture Strategy for the Identification of Nonribosomal Peptide and Polyketide Intermediates.
Article in Methods in molecular biology (Clifton, N.J.), 2023. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Not yet cited in PubMed.
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Abstract
Nonribosomal peptide synthetases (NRPSs) and polyketide synthases (PKSs) are multi-domainal megasynthases. While they are capable of generating a structurally diverse array of metabolites of therapeutic relevance, their mere size and complex nature of their assembly (intermediates are tethered and enzyme bound) make them inherently difficult to characterize. In order to facilitate structural characterization of these metabolites, a thioester capture strategy that enables direct trapping and characterization of the thioester-bound enzyme intermediates was developed. Specifically, a synthetic Biotin-Cys agent was designed and utilized, enabling direct analysis by LCMS/MS and NMR spectroscopy. In the long term, the approach might facilitate the discovery of novel scaffolds from cryptic biosynthetic pathways, paving the way for the development of drug leads and therapeutic initiatives.
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