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Conformational changes in the metallo-beta-lactamase ImiS during the catalytic reaction: an EPR spectrokinetic study of Co(II)-spin label interactions. J Am Chem Soc 2008 Jul 02;130(26):8215-22

Date

06/06/2008

Pubmed ID

18528987

Pubmed Central ID

PMC2574873

DOI

10.1021/ja0774562

Scopus ID

2-s2.0-46049100791 (requires institutional sign-in at Scopus site)   18 Citations

Abstract

Metallo-beta-lactamases are responsible for conferring antibiotic resistance on certain pathogenic bacteria. In consequence, the search for inhibitors that may be useful in combating antibiotic resistance has fueled much study of the active sites of these enzymes. There exists circumstantial evidence that the binding of substrates and inhibitors to metallo-beta-lactamases may involve binding to the organic part of the molecule, in addition to or prior to binding to one or more active site metal ions. It has also been postulated that a conformational change may accompany this putative binding. In the present study, electron paramagnetic resonance spectrokinetic study of a spin-labeled variant of the class B2 metallo-beta-lactamase ImiS identified movement of a component residue on a conserved alpha-helix in a catalytically competent time upon formation of a transient reaction intermediate with the substrate imipenem. In a significant subpopulation of ImiS, this conformational change was not associated with substrate binding to the active site metal ion but, rather, represents a distinct step in the reaction with ImiS. This observation has implications regarding the determinants of substrate specificity in metallo-beta-lactamases and the design of potentially clinically useful inhibitors.

Author List

Sharma N, Hu Z, Crowder MW, Bennett B

Author

Brian Bennett D.Phil. Professor and Chair in the Physics department at Marquette University




MESH terms used to index this publication - Major topics in bold

Bacterial Proteins
Catalysis
Cobalt
Electron Spin Resonance Spectroscopy
Kinetics
Protein Conformation
Spin Labels
Substrate Specificity
beta-Lactamases