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Comparison of Verona Integron-Borne Metallo-β-Lactamase (VIM) Variants Reveals Differences in Stability and Inhibition Profiles

DOI: 10.1128/AAC.01768-15 DOI Help

Authors: Anne Makena (University of Oxford) , Azer Ö. Düzgün (Gümüşhane University) , Jurgen Brem (University of Oxford) , Michael Mcdonough (Department of Chemistry, University of Oxford) , Anna M. Rydzik (University of Oxford) , Martine I. Abboud (University of Oxford) , Ayşegül Saral (Coruh University) , Ayşegül Ç. Çiçek (Recep Tayyip Erdoğan University) , Cemal Sandalli (Recep Tayyip Erdoğan University) , Christopher J. Schofield (University of Oxford)
Co-authored by industrial partner: No

Type: Journal Paper
Journal: Antimicrobial Agents And Chemotherapy , VOL 60 (3) , PAGES 1377 - 1384

State: Published (Approved)
Published: February 2016

Abstract: Metallo-β-lactamases (MBLs) are of increasing clinical significance; the development of clinically useful MBL inhibitors is challenged by the rapid evolution of variant MBLs. The Verona integron-borne metallo-β-lactamase (VIM) enzymes are among the most widely distributed MBLs, with >40 VIM variants having been reported. We report on the crystallographic analysis of VIM-5 and comparison of biochemical and biophysical properties of VIM-1, VIM-2, VIM-4, VIM-5, and VIM-38. Recombinant VIM variants were produced and purified, and their secondary structure and thermal stabilities were investigated by circular dichroism analyses. Steady-state kinetic analyses with a representative panel of β-lactam substrates were carried out to compare the catalytic efficiencies of the VIM variants. Furthermore, a set of metalloenzyme inhibitors were screened to compare their effects on the different VIM variants. The results reveal only small variations in the kinetic parameters of the VIM variants but substantial differences in their thermal stabilities and inhibition profiles. Overall, these results support the proposal that protein stability may be a factor in MBL evolution and highlight the importance of screening MBL variants during inhibitor development programs.

Subject Areas: Biology and Bio-materials


Instruments: I04-Macromolecular Crystallography