An active site loop toggles between conformations to control antibiotic hydrolysis and inhibition potency for CTX-M β-lactamase drug-resistance enzymes

Lu, Shuo; Hu, Liya; Lin, Hanfeng; Judge, Allison; Rivera, Paola; Palaniappan, Murugesan; Sankaran, Banumathi; Wang, Jin; Prasad, B. V. Venkataram; Palzkill, Timothy

doi:10.1038/s41467-022-34564-3

Title: An active site loop toggles between conformations to control antibiotic hydrolysis and inhibition potency for CTX-M β-lactamase drug-resistance enzymes

Journal Article · Mon Nov 07 00:00:00 EST 2022 · Nature Communications

DOI:https://doi.org/10.1038/s41467-022-34564-3· OSTI ID:1897339

Lu, Shuo;

;

; Palaniappan, Murugesan;

;

Abstract β-lactamases inactivate β-lactam antibiotics leading to drug resistance. Consequently, inhibitors of β-lactamases can combat this resistance, and the β-lactamase inhibitory protein (BLIP) is a naturally occurring inhibitor. The widespread CTX-M-14 and CTX-M-15 β-lactamases have an 83% sequence identity. In this study, we show that BLIP weakly inhibits CTX-M-14 but potently inhibits CTX-M-15. The structure of the BLIP/CTX-M-15 complex reveals that binding is associated with a conformational change of an active site loop of β-lactamase. Surprisingly, the loop structure in the complex is similar to that in a drug-resistant variant (N106S) of CTX-M-14. We hypothesized that the pre-established favorable loop conformation of the N106S mutant would facilitate binding. The N106S substitution results in a ~100- and 10-fold increase in BLIP inhibition potency for CTX-M-14 and CTX-M-15, respectively. Thus, this indicates that an active site loop in β-lactamase toggles between conformations that control antibiotic hydrolysis and inhibitor susceptibility. These findings highlight the role of accessible active site conformations in controlling enzyme activity and inhibitor susceptibility as well as the influence of mutations in selectively stabilizing discrete conformations.

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Research Organization:: Lawrence Berkeley National Laboratory (LBNL), Berkeley, CA (United States)

Sponsoring Organization:: National Institutes of Health (NIH); National Institute of General Medical Sciences; USDOE Office of Science (SC); Cancer Prevention & Research Institute of Texas (CPRIT); Welch Foundation

Grant/Contract Number:: AC02-05CH11231; CA259664; AI32956; RP220524; Q1279; T32 GM120011; P30 GM124169-01

OSTI ID:: 1897339

Alternate ID(s):: OSTI ID: 1989647

Journal Information:: Nature Communications, Journal Name: Nature Communications Vol. 13 Journal Issue: 1; ISSN 2041-1723

Publisher:: Nature Publishing GroupCopyright Statement

Country of Publication:: United Kingdom

Language:: English

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59 BASIC BIOLOGICAL SCIENCES
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X-ray crystallography

Title: An active site loop toggles between conformations to control antibiotic hydrolysis and inhibition potency for CTX-M β-lactamase drug-resistance enzymes

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