title: Inhibition of a cold-active alkaline phosphatase by imipenem revealed by in silico modeling of metallo-β-lactamase active sites.
abstract: We demonstrate the inhibition of the native phosphatase activity of a cold active alkaline phosphatase from Vibrio (VAP) (IC(50) of 44±4 (n=4)μM at pH 7.0 after a 30min preincubation) by a specific β-lactam compound (only by imipenem, and not by ertapenem, meropenem, ampicillin or penicillin G). The homologous scaffold was detected by an in silico analysis that established the spatial and electrostatic congruence of the active site of a Class B2 CphA metallo-β-lactamase from Aeromonas hydrophila to the active site of VAP. The tested β-lactam compounds did not inhibit Escherichia coli or shrimp alkaline phosphatase, which could be ascribed to the lower congruence indicated by CLASP. There was no discernible β-lactamase activity in the tested alkaline phosphatases. This is the first time a scaffold recognizing imipenem in an alkaline phosphatase (VAP) has been demonstrated.
authors.affiliation: Department of Biological Sciences, Tata Institute of Fundamental Research, Homi Bhabha Road, Mumbai 400 005, India. sandeepchak@tifr.res.in
authors.forename: Sandeep
authors.initials: S
authors.lastname: Chakraborty
authors.forename: Bjarni
authors.initials: B
authors.lastname: Asgeirsson
authors.forename: Renu
authors.initials: R
authors.lastname: Minda
authors.forename: Lipika
authors.initials: L
authors.lastname: Salaye
authors.forename: Jean-Marie
authors.initials: JM
authors.lastname: Frère
authors.forename: Basuthkar J
authors.initials: BJ
authors.lastname: Rao
mesh_terms: D016980:Aeromonas hydrophila / Q000201:enzymology; D000469:Alkaline Phosphatase / Q000037:antagonists & inhibitors* / Q000737:chemistry / Q000378:metabolism*; D020816:Amino Acid Motifs; D000818:Animals; D020134:Catalytic Domain; D003080:Cold Temperature; D019295:Computational Biology; D004791:Enzyme Inhibitors / Q000494:pharmacology; D006868:Hydrolysis; D015378:Imipenem / Q000494:pharmacology*; D008958:Models, Molecular; D010400:Penicillin G / Q000378:metabolism; D055672:Static Electricity; D001618:beta-Lactamases / Q000737:chemistry* / Q000378:metabolism
chemical_list: D004791:Enzyme Inhibitors; D015378:Imipenem; D000469:Alkaline Phosphatase; D001618:beta-Lactamases; D010400:Penicillin G