title: The three-dimensional structure of VIM-31--a metallo-β-lactamase from Enterobacter cloacae in its native and oxidized form.
abstract: The metallo-β-lactamase VIM-31 differs from VIM-2 by only two Tyr224His and His252Arg substitutions. Located close to the active site, the Tyr224His substitution is also present in VIM-1, VIM-4, VIM-7 and VIM-12. The VIM-31 variant was reported in 2012 from Enterobacter cloacae and kinetically characterized. It exhibits globally lower catalytic efficiencies than VIM-2. In the present study, we report the three-dimensional structures of VIM-31 in its native (reduced) and oxidized forms. The so-called 'flapping-loop' (loop 1) and loop 3 of VIM-31 were not positioned as in VIM-2 but instead were closer to the active site as in VIM-4, resulting in a narrower active site in VIM-31. Also, the presence of His224 in VIM-31 disrupts hydrogen-bonding networks close to the active site. Moreover, a third zinc-binding site, which also exists in VIM-2 structures, could be identified as a structural explanation for the decreased activity of VIM-MBLs at high zinc concentrations.
authors.affiliation: Institute of Molecular Biotechnology, RWTH-Aachen University, Germany.
authors.forename: Michaël B
authors.initials: MB
authors.lastname: Kupper
authors.affiliation: Institute of Molecular Biotechnology, RWTH-Aachen University, Germany.
authors.forename: Konrad
authors.initials: K
authors.lastname: Herzog
authors.affiliation: Institute of Molecular Biotechnology, RWTH-Aachen University, Germany.
authors.forename: Sandra
authors.initials: S
authors.lastname: Bennink
authors.affiliation: Institute of Molecular Biotechnology, RWTH-Aachen University, Germany.
authors.forename: Philipp
authors.initials: P
authors.lastname: Schlömer
authors.affiliation: Laboratory of Bacteriology, CHU Mont-Godinne-Dinant, Université Catholique de Louvain, Yvoir, Belgium.
authors.forename: Pierre
authors.initials: P
authors.lastname: Bogaerts
authors.affiliation: Laboratory of Bacteriology, CHU Mont-Godinne-Dinant, Université Catholique de Louvain, Yvoir, Belgium.
authors.forename: Youri
authors.initials: Y
authors.lastname: Glupczynski
authors.affiliation: Institute of Molecular Biotechnology, RWTH-Aachen University, Germany.
authors.forename: Rainer
authors.initials: R
authors.lastname: Fischer
authors.affiliation: Institute of Molecular Biotechnology, RWTH-Aachen University, Germany.
authors.forename: Carine
authors.initials: C
authors.lastname: Bebrone
authors.affiliation: Institute of Molecular Biotechnology, RWTH-Aachen University, Germany.
authors.forename: Kurt M
authors.initials: KM
authors.lastname: Hoffmann
mesh_terms: D019943:Amino Acid Substitution; D001426:Bacterial Proteins / Q000737:chemistry* / Q000378:metabolism; D001665:Binding Sites; D020134:Catalytic Domain; D016972:Enterobacter cloacae / Q000201:enzymology*; D006860:Hydrogen Bonding; D007527:Isoenzymes / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D007700:Kinetics; D008667:Metalloproteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D010084:Oxidation-Reduction; D011487:Protein Conformation; D017433:Protein Structure, Secondary; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism; D015032:Zinc / Q000378:metabolism; D001618:beta-Lactamases / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism
chemical_list: D001426:Bacterial Proteins; D007527:Isoenzymes; D008667:Metalloproteins; D011994:Recombinant Proteins; D001618:beta-Lactamases; C482459:metallo-beta-lactamase VIM-4, Enterobacter cloacae; D015032:Zinc
keywords: Enterobacter cloacae; active site; antibiotic resistance; crystallographic structures; metallo-β-lactamase