Structural and mechanistic insights into the inhibition of class C beta-lactamases through the adenylylation of the nucleophilic serine
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Title
- Structural and mechanistic insights into the inhibition of class C beta-lactamases through the adenylylation of the nucleophilic serine
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Author(s)
- Kim, Min-Kyu; An, Young Jun; Na, Jung-Hyun; Seol, Jae-Hee; Ryu, Ju Yeon; Lee, Jin-Won; Kang, Lin-Woo; Chung, Kyung Min; Lee, Jung-Hyun; Moon, Jeong Hee; Lee, Jong Seok; Cha, Sun-Shin
- KIOST Author(s)
- An, Young Jun(안영준); Lee, Jung Hyun(이정현); Lee, Jong Seok(이종석)
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Alternative Author(s)
- 안영준; 이정현; 이종석
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Publication Year
- 2017-03
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Abstract
- Objectives: Investigation into the adenylylation of the nucleophilic serine in AmpC BER and CMY-10 extended spectrumclass C beta-lactamases. Methods: The formation and the stability of the adenylate adduct were examined by X-ray crystallography and MS. Inhibition assays for kinetic parameters were performed by monitoring the hydrolytic activity of AmpC BER and CMY-10 using nitrocefin as a reporter substrate. The effect of adenosine 50'-(P-acetyl) monophosphate ( acAMP) on the MIC of ceftazidime was tested with four Gram-negative clinical isolates. Results: The crystal structures and MS analyses confirmed the acAMP-mediated adenylylation of the nucleophilic serine in AmpC BER and CMY-10. acAMP inhibited AmpC BER and CMY-10 through the adenylylation of the nucleophilic serine, which could bemodelled as a two-stepmechanism. The initial non-covalent binding of acAMP to the active site is followed by the covalent attachment of its AMP moiety to the nucleophilic serine. The inhibition efficiencies (k(inact)/K-I) of acAMP against AmpC BER and CMY-10 were determined to be 320 and 140 M(-1)s(-1), respectively. The combination of ceftazidime and acAMP reduced the MIC of ceftazidime against the tested bacteria. Conclusions: Our structural and kinetic studies revealed the detailed mechanismof adenylylation of the nucleophilic serine and may serve as a starting point for the design of novel class C beta-lactamase inhibitors on the basis of the nucleotide scaffold.
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ISSN
- 0305-7453
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URI
- https://sciwatch.kiost.ac.kr/handle/2020.kiost/1279
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DOI
- 10.1093/jac/dkw491
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Bibliographic Citation
- JOURNAL OF ANTIMICROBIAL CHEMOTHERAPY, v.72, no.3, pp.735 - 743, 2017
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Publisher
- OXFORD UNIV PRESS
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Subject
- ENTEROBACTER-CLOACAE P99; EXTENDED-SPECTRUM CEPHALOSPORINASES; TRANSITION-STATE ANALOG; CLASS-A; PHOSPHONATE; ANTIBIOTICS; AVIBACTAM; EPIDEMIOLOGY; INACTIVATION; DERIVATIVES
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Type
- Article
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Language
- English
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Document Type
- Article
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