https://eutils.ncbi.nlm.nih.gov/entrez/eutils/efetch.fcgi?db=pubmed&id=34882867&retmode=xml&tool=Litmetric&email=readroberts32@gmail.com&api_key=61f08fa0b96a73de8c900d749fcb997acc09 348828672022032120230302
1469-896X3132022MarProtein science : a publication of the Protein SocietyProtein SciStructural and functional characterization of fosfomycin resistance conferred by FosB from Enterococcus faecium.580590580-59010.1002/pro.4253The Gram-positive pathogen Enterococcus faecium is one of the leading causes of hospital-acquired vancomycin resistant enterococci (VRE) infections. E. faecium has extensive multidrug resistance and accounts for more than two million infections in the United States each year. FosB is a fosfomycin resistance enzyme found in Gram-positive pathogens like E. faecium. Typically, the FosB enzymes are Mn2+ -dependent bacillithiol (BSH) transferases that inactivate fosfomycin through nucleophilic addition of the thiol to the antibiotic. However, our kinetic analysis of FosBEf shows that the enzyme does not utilize BSH as a thiol substrate, unlike the other well characterized FosB enzymes. Here we report that FosBEf is a Mn2+ -dependent L-cys transferase. In addition, we have determined the three-dimensional X-ray crystal structure of FosBEf in complex with fosfomycin at a resolution of 2.0 Å. A sequence similarity network (SSN) was generated for the FosB family to investigate the unexpected substrate selectivity. Three non-conserved residues were identified in the SSN that may contribute to the substrate selectivity differences in the family of enzymes. Our structural and functional characterization of FosBEf establishes the enzyme as a potential target and may prove useful for future structure-based development of FosB inhibitors to increase the efficacy of fosfomycin.© 2021 The Protein Society.WiltsieVanessaVDepartment of Chemistry & Biochemistry, The University of Alabama, Tuscaloosa, Alabama, USA.TravisSkyeSDepartment of Chemistry & Biochemistry, The University of Alabama, Tuscaloosa, Alabama, USA.ShayMadeline RMRDepartment of Chemistry & Biochemistry, The University of Alabama, Tuscaloosa, Alabama, USA.SimmonsZacharyZDepartment of Chemistry & Biochemistry, The University of Alabama, Tuscaloosa, Alabama, USA.FrantomPatrickPDepartment of Chemistry & Biochemistry, The University of Alabama, Tuscaloosa, Alabama, USA.ThompsonMatthew KMK0000-0002-1964-3896Department of Chemistry & Biochemistry, The University of Alabama, Tuscaloosa, Alabama, USA.engJournal ArticleResearch Support, N.I.H., ExtramuralResearch Support, Non-U.S. Gov't20211222
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