Development of a selective activity-based probe for adenylating enzymes: profiling MbtA Involved in siderophore biosynthesis from Mycobacterium tuberculosis.
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.
Center for Drug Design and ‡Department of Medicinal Chemistry, University of Minnesota, Minneapolis, Minnesota 55455, United States.
Benjamin P
BP
Duckworth
Daniel J
DJ
Wilson
Kathryn M
KM
Nelson
Helena I
HI
Boshoff
Clifton E
CE
Barry
Courtney C
CC
Aldrich
D000225:Adenine / Q000378:metabolism*; D000241:Adenosine / Q000031:analogs & derivatives / Q000494:pharmacology; D000995:Antitubercular Agents / Q000494:pharmacology; D008025:Ligases / Q000037:antagonists & inhibitors* / Q000302:isolation & purification; D008826:Microbial Sensitivity Tests; D008958:Models, Molecular; D009169:Mycobacterium tuberculosis / Q000201:enzymology* / Q000378:metabolism*; D017262:Siderophores / Q000378:metabolism*; D013329:Structure-Activity Relationship
C526816:5'-O-(N-(salicyl)sulfamoyl)adenosine; D000995:Antitubercular Agents; D017262:Siderophores; D008025:Ligases; C526815:salicyl-AMP ligase, Mycobacterium tuberculosis; D000225:Adenine; D000241:Adenosine