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Title: Analysis of the Structure and Function of FOX-4 Cephamycinase

Journal Article · · Antimicrobial Agents and Chemotherapy
DOI:https://doi.org/10.1128/aac.01887-15· OSTI ID:1243116
 [1];  [2];  [2];  [2];  [1];  [1];  [1];  [2];  [3];  [4];  [2];  [5];  [6];  [4]
  1. Hofstra Univ., Hempstead, NY (United States)
  2. Albert Einstein College of Medicine, Bronx, NY (United States)
  3. Case Western Reserve Univ., Cleveland, OH (United States); Université de Liège (Belgium)
  4. Case Western Reserve Univ., Cleveland, OH (United States)
  5. Université de Liège (Belgium)
  6. Universitario A. Coruña, La Coruña (Spain)
+ Show Author Affiliations

Class C β-lactamases poorly hydrolyze cephamycins (e.g., cefoxitin, cefotetan, and moxalactam). In the past 2 decades, a new family of plasmid-based AmpC β-lactamases conferring resistance to cefoxitin, the FOX family, has grown to include nine unique members descended from the Aeromonas caviae chromosomal AmpC. To understand the basis for the unique cephamycinase activity in the FOX family, we determined the first X-ray crystal structures of FOX-4, apo enzyme and the acyl-enzyme with its namesake compound, cefoxitin, using the Y150F deacylation-deficient variant. Notably, recombinant expression of N-terminally tagged FOX-4 also yielded an inactive adenylylated enzyme form not previously observed in β-lactamases. The posttranslational modification (PTM), which occurs on the active site Ser64, would not seem to provide a selective advantage, yet might present an opportunity for the design of novel antibacterial drugs. Substantial ligand-induced changes in the enzyme are seen in the acyl-enzyme complex, particularly the R2 loop and helix H10 (P289 to N297), with movement of F293 by 10.3 Å. Taken together, this study provides the first picture of this highly proficient class C cephamycinase, uncovers a novel PTM, and suggests a possible cephamycin resistance mechanism involving repositioning of the substrate due to the presence of S153P, N289P, and N346I substitutions in the ligand binding pocket.

Research Organization:
Argonne National Laboratory (ANL), Argonne, IL (United States)
Sponsoring Organization:
USDOE Office of Science (SC)
Grant/Contract Number:
AC02- 06CH11357
OSTI ID:
1243116
Journal Information:
Antimicrobial Agents and Chemotherapy, Vol. 60, Issue 2; ISSN 0066-4804
Publisher:
American Society for MicrobiologyCopyright Statement
Country of Publication:
United States
Language:
ENGLISH
Citation Metrics:
Cited by: 14 works
Citation information provided by
Web of Science

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Cited By (4)

Atomic-Resolution Structure of a Class C β-Lactamase and Its Complex with Avibactam journal June 2018
Probing the Mechanism of Inactivation of the FOX-4 Cephamycinase by Avibactam journal February 2018
Emergence of Resistance to Novel β-Lactam–β-Lactamase Inhibitor Combinations Due to Horizontally Acquired AmpC (FOX-4) in Pseudomonas aeruginosa Sequence Type 308 journal November 2019
In vitro and in vivo Inhibitory Activity of NADPH Against the AmpC BER Class C β-Lactamase journal December 2018

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