Structural and mutagenic analysis of metallo-β-lactamase IMP-18

Takamitsu Furuyama, Haruka Nonomura, Yoshikazu Ishii, Nancy D. Hanson, Akiko Shimizu-Ibuka

Research output: Contribution to journalArticle

2 Citations (Scopus)

Abstract

IMP-type metallo-β-lactamases (MBLs) are exogenous zinc metalloenzymes that hydrolyze a broad range of β-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 αβ/βα "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 β-lactamases.

Original languageEnglish (US)
Pages (from-to)5521-5526
Number of pages6
JournalAntimicrobial Agents and Chemotherapy
Volume60
Issue number9
DOIs
StatePublished - Sep 1 2016

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Inosine Monophosphate
meropenem
Lactams
Carbapenems
Pseudomonas aeruginosa
Zinc
Catalytic Domain
Amino Acids

All Science Journal Classification (ASJC) codes

  • Pharmacology
  • Infectious Diseases
  • Pharmacology (medical)

Cite this

Structural and mutagenic analysis of metallo-β-lactamase IMP-18. / Furuyama, Takamitsu; Nonomura, Haruka; Ishii, Yoshikazu; Hanson, Nancy D.; Shimizu-Ibuka, Akiko.

In: Antimicrobial Agents and Chemotherapy, Vol. 60, No. 9, 01.09.2016, p. 5521-5526.

Research output: Contribution to journalArticle

Furuyama, Takamitsu ; Nonomura, Haruka ; Ishii, Yoshikazu ; Hanson, Nancy D. ; Shimizu-Ibuka, Akiko. / Structural and mutagenic analysis of metallo-β-lactamase IMP-18. In: Antimicrobial Agents and Chemotherapy. 2016 ; Vol. 60, No. 9. pp. 5521-5526.
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