Structural insights into the inhibition of the extended-spectrum -lactamase PER-2 by avibactam
American Society for Microbiology, 2019
Online
unknown
Zugriff:
The diazabicyclooctane (DBO) avibactam (AVI) reversibly inactivates most serine-lactamases. Previous investigations showed that inhibition constants of AVI toward class A PER-2 are reminiscent of values observed for class C and D -lactamases (i.e., k2/K of 103 M1 s1) but lower than other class A -lactamases (i.e., k2/K 104 to 105 M1 s1). Herein, biochemical and structural studies were conducted with PER-2 and AVI to explore these differences. Furthermore, biochemical studies on Arg220 and Thr237 variants with AVI were conducted to gain deeper insight into the mechanism of PER-2 inactivation. The main biochemical and structural observations revealed the following: (i) both amino-acid substitutions in Arg220 and the rich hydrophobic content in the active site hinder the binding of catalytic waters and acylation, impairing AVI inhibition; (ii) movement of Ser130 upon binding of AVI favors the formation of a hydrogen bond with the sulfate group of AVI; and (iii) the Thr237Ala substitution alters the AVI inhibition constants. The acylation constant (k2/K) of PER-2 by AVI is primarily influenced by stabilizing hydrogen bonds involving AVI and important residues such as Thr237 and Arg220. (Variants in Arg220 demonstrate a dramatic reduction in k2/K.) We also observed that displacement of Ser130 side chain impairs AVI acylation, an observation not made in other extended-spectrum -lactamases (ESBLs). Comparatively, relebactam combined with a -lactam is more potent against Escherichia coli producing PER-2 variants than -lactam–AVI combinations. Our findings provide a rationale for evaluating the utility of the currently available DBO inhibitors against unique ESBLs like PER-2 and anticipate the effectiveness of these inhibitors toward variants that may eventually be selected upon AVI usage. Fil: Ruggiero, Melina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Departamento de Microbiología, Inmunología y Biotecnología. Cátedra de Microbiología; Argentina Fil: Papp Wallace, Krisztina M.. Case Western Reserve University. School of Medicine; Estados Unidos. Louis Stokes Cleveland VA Medical Center; Estados Unidos Fil: Brunetti, Florencia Lourdes. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Departamento de Microbiología, Inmunología y Biotecnología. Cátedra de Microbiología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Barnes, Melissa D.. Case Western Reserve University. School of Medicine; Estados Unidos. Louis Stokes Cleveland VA Medical Center; Estados Unidos Fil: Bonomo, Robert A.. Louis Stokes Cleveland VA Medical Center; Estados Unidos. Case Western Reserve University. School of Medicine; Estados Unidos. Center for Antimicrobial Resistance and Epidemiology; Estados Unidos Fil: Gutkind, Gabriel Osvaldo. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Departamento de Microbiología, Inmunología y Biotecnología. Cátedra de Microbiología; Argentina Fil: Klinke, Sebastian. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Investigaciones Bioquímicas de Buenos Aires. Fundación Instituto Leloir. Instituto de Investigaciones Bioquímicas de Buenos Aires; Argentina. Plataforma Argentina de Biología Estructural y Metabolómica; Argentina Fil: Power, Pablo. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Departamento de Microbiología, Inmunología y Biotecnología. Cátedra de Microbiología; Argentina
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Structural insights into the inhibition of the extended-spectrum -lactamase PER-2 by avibactam
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Autor/in / Beteiligte Person: | Brunetti, Florencia ; Gabriel Osvaldo Gutkind ; Ruggiero, Melina ; Papp-Wallace, Krisztina M. ; Barnes, Melissa D. ; Klinke, Sebastián ; Bonomo, Robert A. ; Power, Pablo |
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Veröffentlichung: | American Society for Microbiology, 2019 |
Medientyp: | unknown |
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