Phosphate Promotes the Recovery of Mycobacterium tuberculosis β‑Lactamase from Clavulanic Acid Inhibition

The rise of multi- and even totally antibiotic resistant forms of Mycobacterium tuberculosis underlines the need for new antibiotics. The pathogen is resistant to β-lactam compounds due to its native serine β-lactamase, BlaC. This resistance can be circumvented by administration of a β-lactamase inh...

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Published in:	Biochemistry (Easton) Vol. 56; no. 47; pp. 6257 - 6267
Main Authors:	Elings, Wouter, Tassoni, Raffaella, van der Schoot, Steven A, Luu, Wendy, Kynast, Josef P, Dai, Lin, Blok, Anneloes J, Timmer, Monika, Florea, Bogdan I, Pannu, Navraj S, Ubbink, Marcellus
Format:	Journal Article
Language:	English
Published:	United States American Chemical Society 28-11-2017
Subjects:	beta-Lactamase Inhibitors - pharmacology beta-Lactamases - chemistry Binding Sites Clavulanic Acid - pharmacology Drug Resistance, Bacterial - drug effects Drug Synergism Humans Hydrolysis Microbial Sensitivity Tests Models, Molecular Mycobacterium tuberculosis - drug effects Phosphates - pharmacology Tuberculosis - drug therapy Tuberculosis - microbiology
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Summary:	The rise of multi- and even totally antibiotic resistant forms of Mycobacterium tuberculosis underlines the need for new antibiotics. The pathogen is resistant to β-lactam compounds due to its native serine β-lactamase, BlaC. This resistance can be circumvented by administration of a β-lactamase inhibitor. We studied the interaction between BlaC and the inhibitor clavulanic acid. Our data show hydrolysis of clavulanic acid and recovery of BlaC activity upon prolonged incubation. The rate of clavulanic acid hydrolysis is much higher in the presence of phosphate ions. A specific binding site for phosphate is identified in the active site pocket, both in the crystalline state and in solution. NMR spectroscopy experiments show that phosphate binds to this site with a dissociation constant of 30 mM in the free enzyme. We conclude that inhibition of BlaC by clavulanic acid is reversible and that phosphate ions can promote the hydrolysis of the inhibitor.
Bibliography:	ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 23
ISSN:	0006-2960 1520-4995
DOI:	10.1021/acs.biochem.7b00556