Development of a selective activity-based probe for adenylating enzymes: Profiling MbtA involved in siderophore biosynthesis from mycobacterium tuberculosis

Benjamin P. Duckworth, Daniel J. Wilson, Kathryn M. Nelson, Helena I. Boshoff, Clifton E. Barry, Courtney C. Aldrich

Research output: Contribution to journalArticlepeer-review

34 Scopus citations

Abstract

MbtA is an adenylating enzyme from Mycobacterium tuberculosis that catalyzes the first step in the biosynthesis of the mycobactins. A bisubstrate inhibitor of MbtA (Sal-AMS) was previously described that displays potent antitubercular activity under iron-replete as well as iron-deficient growth conditions. This finding is surprising since mycobactin biosynthesis is not required under iron-replete conditions and suggests off-target inhibition of additional biochemical pathways. As a first step toward a complete understanding of the mechanism of action of Sal-AMS, we have designed and validated an activity-based probe (ABP) for studying Sal-AMS inhibition in M. tuberculosis. This probe labels pure MbtA as well as MbtA in mycobacterial lysate, and labeling can be completely inhibited by preincubation with Sal-AMS. Furthermore, this probe provides a prototypical core scaffold for the creation of ABPs to profile any of the other 66 adenylating enzymes in Mtb or the multitude of adenylating enzymes in other pathogenic bacteria.

Original languageEnglish (US)
Pages (from-to)1653-1658
Number of pages6
JournalACS Chemical Biology
Volume7
Issue number10
DOIs
StatePublished - Oct 19 2012

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