期刊
ANTIMICROBIAL AGENTS AND CHEMOTHERAPY
卷 56, 期 5, 页码 2713-2718出版社
AMER SOC MICROBIOLOGY
DOI: 10.1128/AAC.06099-11
关键词
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资金
- Robert A. Welch Foundation [N-0871]
- National Institutes of Health [R01 AI062968, RO1 AI063517-01]
- Veterans Affairs Merit Review Program
- Geriatric Research Education and Clinical Care (GRECC)
- Veterans Affairs Career Development Program
Class A carbapenemases are a major threat to the potency of carbapenem antibiotics. A widespread carbapenemase, KPC-2, is not easily inhibited by beta-lactamase inhibitors (i.e., clavulanic acid, sulbactam, and tazobactam). To explore different mechanisms of inhibition of KPC-2, we determined the crystal structures of KPC-2 with two beta-lactamase inhibitors that follow different inactivation pathways and kinetics. The first complex is that of a small boronic acid compound, 3-nitrophenyl boronic acid (3-NPBA), bound to KPC-2 with 1.62-angstrom resolution. 3-NPBA demonstrated a K-m value of 1.0 +/- 0.1 mu M (mean +/- standard error) for KPC-2 and blocks the active site by making a reversible covalent interaction with the catalytic S70 residue. The two boron hydroxyl atoms of 3-NPBA are positioned in the oxyanion hole and the deacylation water pocket, respectively. In addition, the aromatic ring of 3-NPBA provides an edge-to-face interaction with W105 in the active site. The structure of KPC-2 with the penam sulfone PSR-3-226 was determined at 1.26-angstrom resolution. PSR-3-226 displayed a K-m value of 3.8 +/- 0.4 mu M for KPC-2, and the inactivation rate constant (k(inact)) was 0.034 +/- 0.003 s(-1). When covalently bound to S70, PSR-3-226 forms a trans-enamine intermediate in the KPC-2 active site. The predominant active site interactions are generated via the carbonyl oxygen, which resides in the oxyanion hole, and the carboxyl moiety of PSR-3-226, which interacts with N132, N170, and E166. 3-NPBA and PSR-3-226 are the first beta-lactamase inhibitors to be trapped as an acyl-enzyme complex with KPC-2. The structural and inhibitory insights gained here could aid in the design of potent KPC-2 inhibitors.
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