title: Kinetic Studies on CphA Mutants Reveal the Role of the P158-P172 Loop in Activity versus Carbapenems.
abstract: Site-directed mutagenesis of CphA indicated that prolines in the P158-P172 loop are essential for the stability and the catalytic activity of subclass B2 metallo-β-lactamases against carbapenems. The sequential substitution of proline led to a decrease of the catalytic efficiency of the variant compared to the wild-type (WT) enzyme but also to a higher affinity for the binding of the second zinc ion.
authors.affiliation: Dipartimento di Scienze Cliniche Applicate e Biotecnologiche, Università degli Studi dell'Aquila, L'Aquila, Italy.
authors.forename: Carlo
authors.initials: C
authors.lastname: Bottoni
authors.affiliation: Dipartimento di Scienze Cliniche Applicate e Biotecnologiche, Università degli Studi dell'Aquila, L'Aquila, Italy.
authors.forename: Mariagrazia
authors.initials: M
authors.lastname: Perilli
authors.affiliation: Dipartimento di Scienze Cliniche Applicate e Biotecnologiche, Università degli Studi dell'Aquila, L'Aquila, Italy.
authors.forename: Francesca
authors.initials: F
authors.lastname: Marcoccia
authors.affiliation: Dipartimento di Scienze Cliniche Applicate e Biotecnologiche, Università degli Studi dell'Aquila, L'Aquila, Italy.
authors.forename: Alessandra
authors.initials: A
authors.lastname: Piccirilli
authors.affiliation: Dipartimento di Scienze Cliniche Applicate e Biotecnologiche, Università degli Studi dell'Aquila, L'Aquila, Italy.
authors.forename: Cristina
authors.initials: C
authors.lastname: Pellegrini
authors.affiliation: Dipartimento di Scienze Cliniche Applicate e Biotecnologiche, Università degli Studi dell'Aquila, L'Aquila, Italy.
authors.forename: Martina
authors.initials: M
authors.lastname: Colapietro
authors.affiliation: Dipartimento di Scienze Cliniche Applicate e Biotecnologiche, Università degli Studi dell'Aquila, L'Aquila, Italy.
authors.forename: Alessia
authors.initials: A
authors.lastname: Sabatini
authors.affiliation: Dipartimento di Scienze Cliniche Applicate e Biotecnologiche, Università degli Studi dell'Aquila, L'Aquila, Italy.
authors.forename: Giuseppe
authors.initials: G
authors.lastname: Celenza
authors.affiliation: Macromolécules Biologiques, Centre d'Ingénierie des Protéines (CIP), Université de Liège, Institut de Chimie B6, Sart Tilman, Liège, Belgium.
authors.forename: Frédéric
authors.initials: F
authors.lastname: Kerff
authors.affiliation: Dipartimento di Scienze Cliniche Applicate e Biotecnologiche, Università degli Studi dell'Aquila, L'Aquila, Italy.
authors.forename: Gianfranco
authors.initials: G
authors.lastname: Amicosante
authors.affiliation: Macromolécules Biologiques, Centre d'Ingénierie des Protéines (CIP), Université de Liège, Institut de Chimie B6, Sart Tilman, Liège, Belgium.
authors.forename: Moreno
authors.initials: M
authors.lastname: Galleni
authors.affiliation: Macromolécules Biologiques, Centre d'Ingénierie des Protéines (CIP), Université de Liège, Institut de Chimie B6, Sart Tilman, Liège, Belgium pmercuri@ulg.ac.be.
authors.forename: Paola Sandra
authors.initials: PS
authors.lastname: Mercuri
mesh_terms: D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism*; D001665:Binding Sites; D015780:Carbapenems / Q000494:pharmacology*; D007700:Kinetics; D008958:Models, Molecular; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D011392:Proline / Q000737:chemistry / Q000378:metabolism; D020539:Sequence Analysis, Protein; D013329:Structure-Activity Relationship; D013379:Substrate Specificity / Q000235:genetics / Q000502:physiology; D015032:Zinc / Q000494:pharmacology; D001618:beta-Lactamases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*
chemical_list: D001426:Bacterial Proteins; D015780:Carbapenems; D011392:Proline; D001618:beta-Lactamases; D015032:Zinc