Please use this identifier to cite or link to this item:
Type: Artigo de periódico
Title: The zn2 position in metallo-beta-lactamases is critical for activity: A study on chimeric metal sites on a conserved protein scaffold
Author: Gonzalez, JM
Martin, FJM
Costello, AL
Tierney, DL
Vila, AJ
Abstract: Metallo beta-lactamases (M beta Ls)are bacterial Zn(II)-dependent hydrolases that, confer broad-spectrum resistance to beta-lactam antibiotics. These enzymes can be subdivided into three subclasses (B1, B2 and B3) that differ in their metal binding sites and their characteristic tertiary structure. To date there are no clinically useful pan-M beta L inhibitors available, mainly due to the unawareness of key catalytic features common to all M beta L brands. Here we have designed, expressed and characterized two double mutants of BcII, a di-Zn(II) B1-M beta L from Bacillus cereus, namely BcII-R121H/C221D (BcII-HD) and BcII-R121H/C221S (BcII-HS). These mutants display modified environments at the so-called Zn2 site or DCH site, reproducing the metal coordination environments of structurally related metallohydrolases. Through a combination of structural and functional studies, we found that BcII-HD is an impaired beta-lactamase even as a di-Zn(II) enzyme, whereas BcII-HS exhibits the ability to exist as mono or di-Zn(II) species in solution, with different catalytic performances. We show that these effects result from an altered position of Zn2, which is incapable of providing a productive interaction with the substrate beta-lactam ring. These results indicate that the position of Zn2 is essential for a productive substrate binding and hydrolysis. (C) 2007 Elsevier Ltd. All rights reserved.
Subject: antibiotic resistance
Zn2 site
catalytic mechanism
Country: Inglaterra
Editor: Academic Press Ltd Elsevier Science Ltd
Rights: fechado
Identifier DOI: 10.1016/j.jmb.2007.08.031
Date Issue: 2007
Appears in Collections:Unicamp - Artigos e Outros Documentos

Files in This Item:
File Description SizeFormat 
WOS000250712600005.pdf1.32 MBAdobe PDFView/Open

Items in DSpace are protected by copyright, with all rights reserved, unless otherwise indicated.