Bottoni Carlo, Perilli Mariagrazia, Marcoccia Francesca, Piccirilli Alessandra, Pellegrini Cristina, Colapietro Martina, Sabatini Alessia, Celenza Giuseppe, Kerff Frédéric, Amicosante Gianfranco, Galleni Moreno, Mercuri Paola Sandra
Dipartimento di Scienze Cliniche Applicate e Biotecnologiche, Università degli Studi dell'Aquila, L'Aquila, Italy.
Macromolécules Biologiques, Centre d'Ingénierie des Protéines (CIP), Université de Liège, Institut de Chimie B6, Sart Tilman, Liège, Belgium.
Antimicrob Agents Chemother. 2016 Apr 22;60(5):3123-6. doi: 10.1128/AAC.01703-15. Print 2016 May.
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.
CphA的定点诱变表明,P158 - P172环中的脯氨酸对于B2亚类金属β-内酰胺酶对碳青霉烯类的稳定性和催化活性至关重要。与野生型(WT)酶相比,脯氨酸的顺序取代导致变体的催化效率降低,但也导致对第二个锌离子结合的更高亲和力。