title: Identification and characterization of an unusual metallo-β-lactamase from Serratia proteamaculans.
abstract: Metallo-β-lactamases (MBLs) are a family of metalloenzymes that are capable of hydrolyzing β-lactam antibiotics and are an important means by which bacterial pathogens use to inactivate antibiotics. A database search of the available amino acid sequences from Serratia proteamaculans indicates the presence of an unusual MBL. A full length amino acid sequence alignment indicates overall homology to B3-type MBLs, but also suggests considerable variations in the active site, notably among residues that are relevant to metal ion binding. Steady-state kinetic measurements further indicate functional differences and identify two relevant pK a values for catalysis (3.8 for the enzyme-substrate complex and 7.8 for the free enzyme) and a preference for penams with modest reactivity towards some cephalosporins. An analysis of the metal ion content indicates the presence of only one zinc ion per active site in the resting enzyme. In contrast, kinetic data suggest that the enzyme may operate as a binuclear enzyme, and it is thus proposed that a catalytically active di-Zn(2+) center is formed only once the substrate is present.
authors: School of Chemistry and Molecular Biosciences, The University of Queensland, St. Lucia, QLD, 4072, Australia.
authors: Peter
authors: P
authors: Vella
authors: Manfredi
authors: M
authors: Miraula
authors: Emer
authors: E
authors: Phelan
authors: Eleanor W W
authors: EW
authors: Leung
authors: Fernanda
authors: F
authors: Ely
authors: David L
authors: DL
authors: Ollis
authors: Ross P
authors: RP
authors: McGeary
authors: Gerhard
authors: G
authors: Schenk
authors: Nataša
authors: N
authors: Mitić
mesh_terms: D000595:Amino Acid Sequence; D055162:Biocatalysis; D030562:Databases, Protein; D055808:Drug Discovery; D008670:Metals; D008958:Models, Molecular; D008969:Molecular Sequence Data; D055503:Protein Multimerization; D020836:Protein Structure, Quaternary; D012705:Serratia / Q000201:enzymology*; D001618:beta-Lactamases / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*
chemical_list: D008670:Metals; D001618:beta-Lactamases
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