Difference in the Inhibitory Effect of Thiol Compounds and Demetallation Rates from the Zn(II) Active Site of Metallo-β-lactamases (IMP-1 and IMP-6) Associated with a Single Amino Acid Substitution.

Autor: Yamaguchi Y; Environmental Safety Center, Kumamoto University, 39-1 Kurokami 2-Chome, Chuo-ku, Kumamoto860-8555, Japan.; Graduate School of Science and Technology, Kumamoto University, 39-1 Kurokami 2-Chome, Chuo-ku, Kumamoto860-8555, Japan.; Faculty of Engineering, Kumamoto University, 39-1 Kurokami 2-Chome, Chuo-ku, Kumamoto860-8555, Japan., Kato K; College of Pharmacy, Kinjo Gakuin University, 2-1723 Omori, Moriyama-ku, Nagoya, Aichi463-8521, Japan.; Faculty of Pharmacy, Meijo University, 150 Yagotoyama, Tempaku-ku, Nagoya, Aichi468-8503, Japan.; Faculty of Pharmaceutical Sciences, Shonan University of Medical Sciences, 16-48, Kamishinano, Totsuka-ku, Yokohama, Kanagawa244-0806, Japan., Ichimaru Y; College of Pharmacy, Kinjo Gakuin University, 2-1723 Omori, Moriyama-ku, Nagoya, Aichi463-8521, Japan.; Faculty of Pharmaceutical Sciences, Shonan University of Medical Sciences, 16-48, Kamishinano, Totsuka-ku, Yokohama, Kanagawa244-0806, Japan., Uenosono Y; Graduate School of Science and Technology, Kumamoto University, 39-1 Kurokami 2-Chome, Chuo-ku, Kumamoto860-8555, Japan., Tawara S; Graduate School of Science and Technology, Kumamoto University, 39-1 Kurokami 2-Chome, Chuo-ku, Kumamoto860-8555, Japan., Ito R; Graduate School of Science and Technology, Kumamoto University, 39-1 Kurokami 2-Chome, Chuo-ku, Kumamoto860-8555, Japan., Matsuse N; Faculty of Engineering, Kumamoto University, 39-1 Kurokami 2-Chome, Chuo-ku, Kumamoto860-8555, Japan., Wachino JI; Department of Medical Technology, Faculty of Medical Sciences, Shubun University, 6 Nikko-cho, Ichinomiya, Aichi491-0938, Japan., Toma-Fukai S; Graduate School of Science and Technology, Nara Institute of Science and Technology, 8916-5 Takayama-cho, Ikoma, Nara630-0192, Japan., Jin W; College of Pharmacy, Kinjo Gakuin University, 2-1723 Omori, Moriyama-ku, Nagoya, Aichi463-8521, Japan., Arakawa Y; Department of Bacteriology, Nagoya University Graduate School of Medicine, 65 Tsurumai-cho, Showa-ku, Nagoya, Aichi466-8550, Japan., Otsuka M; Medicinal and Biological Chemistry Science Farm Joint Research Laboratory, Faculty of Life Sciences, Kumamoto University, 5-1 Oe-honmachi, Chuo-ku, Kumamoto862-0973, Japan.; Department of Drug Discovery, Science Farm Ltd., 1-7-30 Kuhonji, Chuo-ku, Kumamoto862-0976, Japan., Fujita M; Medicinal and Biological Chemistry Science Farm Joint Research Laboratory, Faculty of Life Sciences, Kumamoto University, 5-1 Oe-honmachi, Chuo-ku, Kumamoto862-0973, Japan., Fukuishi N; College of Pharmacy, Kinjo Gakuin University, 2-1723 Omori, Moriyama-ku, Nagoya, Aichi463-8521, Japan., Sugiura K; College of Pharmacy, Kinjo Gakuin University, 2-1723 Omori, Moriyama-ku, Nagoya, Aichi463-8521, Japan., Imai M; College of Pharmacy, Kinjo Gakuin University, 2-1723 Omori, Moriyama-ku, Nagoya, Aichi463-8521, Japan., Kurosaki H; College of Pharmacy, Kinjo Gakuin University, 2-1723 Omori, Moriyama-ku, Nagoya, Aichi463-8521, Japan.
Jazyk: angličtina
Zdroj: ACS infectious diseases [ACS Infect Dis] 2023 Jan 13; Vol. 9 (1), pp. 65-78. Date of Electronic Publication: 2022 Dec 15.
DOI: 10.1021/acsinfecdis.2c00395
Abstrakt: Gram-negative bacteria producing metallo-β-lactamases (MBLs) have become a considerable threat to public health. MBLs including the IMP, VIM, and NDM types are Zn(II) enzymes that hydrolyze the β-lactam ring present in a broad range of antibiotics, such as N -benzylpenicillin, meropenem, and imipenem. Among IMPs, IMP-1 and IMP-6 differ in a single amino acid substitution at position 262, where serine in IMP-1 is replaced by glycine in IMP-6, conferring a change in substrate specificity. To investigate how this mutation influences enzyme function, we examined lactamase inhibition by thiol compounds. Ethyl 3-mercaptopropionate acted as a competitive inhibitor of IMP-1, but a noncompetitive inhibitor of IMP-6. A comparison of the crystal structures previously reported for IMP-1 (PDB code: 5EV6) and IMP-6 (PDB code: 6LVJ) revealed a hydrogen bond between the side chain of Ser262 and Cys221 in IMP-1 but the absence of hydrogen bond in IMP-6, which affects the Zn2 coordination sphere in its active site. We investigated the demetallation rates of IMP-1 and IMP-6 in the presence of chelating agent ethylenediaminetetraacetic acid (EDTA) and found that the demetallation reactions had fast and slow phases with a first-order rate constant ( k fast = 1.76 h -1 , k slow = 0.108 h -1 for IMP-1, and k fast = 14.0 h -1 and k slow = 1.66 h -1 for IMP-6). The difference in the flexibility of the Zn2 coordination sphere between IMP-1 and IMP-6 may influence the demetallation rate, the catalytic efficiency against β-lactam antibiotics, and the inhibitory effect of thiol compounds.
Databáze: MEDLINE
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