Structural and Mutagenic Analysis of Metallo-?-Lactamase IMP-18.

ABSTRACT: IMP-type metallo-?-lactamases (MBLs) are exogenous zinc metalloenzymes that hydrolyze a broad range of ?-lactams, including carbapenems. Here we report the crystal structure of IMP-18, an MBL cloned from Pseudomonas aeruginosa, at 2.0-Å resolution. The overall structure of IMP-18 resembles that of IMP-1, with an ??/?? "folded sandwich" configuration, but the loop that covers the active site has a distinct conformation. The relationship between IMP-18's loop conformation and its kinetic properties was investigated by replacing the amino acid residues that can affect the loop conformation (Lys44, Thr50, and Ile69) in IMP-18 with those occupying the corresponding positions in the well-described enzyme IMP-1. The replacement of Thr50 with Pro considerably modified IMP-18's kinetic properties, specifically those pertaining to meropenem, with the kcat/Km value increased by an order of magnitude. The results indicate that this is a key residue that defines the kinetic properties of IMP-type ?-lactamases.

SUBMITTER: Furuyama T 

PROVIDER: S-EPMC4997881 | biostudies-literature | 2016 Sep

REPOSITORIES: biostudies-literature