Multi-drug resistant Gram negative bacteria (MDR-GNB) present a significant threat to successful antimicrobial chemotherapy. Among MDR-GNB, extended spectrum beta-lactamase (ESBL) producing Klebsiella pneumoniae resistant to ceftazidime are associated with increased mortality. The atomic structure determination of the SHV-2 beta-lactamase, the first ESBL described, revealed the active site of the enzyme is widened to accommodate the side chain of ceftazidime. Despite this insight, our understanding is still incomplete. The broad, long-term objectives of this proposal are: 1) To determine the atomic structures of boronic acid transition-state analogue inhibitors that have the R1 side chains of ceftazidime and cefotaxime in the SHV-1, -2 and -5 beta-lactamases. This will elucidate the critical interactions that define the ability of ESBLs to hydrolyze advanced generation cephalosporins. 2) Using strains of E. coli deficient in the principal DNA mismatch repair protein, mutS, select blaSHV mutants that stabilize and enhance expression of SHV beta-lactamase. 3) To determine if mutator phenotypes exist in K. pneumoniae;assess the relationship between ESBL enzymes, mutator phenotypes, and mutS expression. 4) To develop a model for ESBL evolution using blaSHV in a mutS deficient strain of E. coli. Accomplishing these goals will achieve an unprecedented understanding of the protein and substrate interactions and genetic properties responsible for the ESBL phenotype. This work will also serve as a paradigm for the prediction of novel ESBL phenotypes in SHV enzymes, form a basis for the evaluation of novel beta-lactams, and show how enzyme drug targets influence substrate affinity and catalysis.

National Institute of Health (NIH)
National Institute of Allergy and Infectious Diseases (NIAID)
Research Project (R01)
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Special Emphasis Panel (ZRG1-IDM-N (90))
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Huntley, Clayton C
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Case Western Reserve University
Internal Medicine/Medicine
Schools of Medicine
United States
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El Chakhtoura, Nadim G; Saade, Elie; Iovleva, Alina et al. (2018) Therapies for multidrug resistant and extensively drug-resistant non-fermenting gram-negative bacteria causing nosocomial infections: a perilous journey toward 'molecularly targeted' therapy. Expert Rev Anti Infect Ther 16:89-110
Nukaga, Michiyoshi; Papp-Wallace, Krisztina M; Hoshino, Tyuji et al. (2018) Probing the Mechanism of Inactivation of the FOX-4 Cephamycinase by Avibactam. Antimicrob Agents Chemother 62:
Kanwar, Anubhav; Marshall, Steven H; Perez, Federico et al. (2018) Emergence of Resistance to Colistin During the Treatment of Bloodstream Infection Caused by Klebsiella pneumoniae Carbapenemase-Producing Klebsiella pneumoniae. Open Forum Infect Dis 5:ofy054
Ghiglione, Barbara; Rodríguez, María Margarita; Curto, Lucrecia et al. (2018) Defining Substrate Specificity in the CTX-M Family: the Role of Asp240 in Ceftazidime Hydrolysis. Antimicrob Agents Chemother 62:
Papp-Wallace, Krisztina M; Barnes, Melissa D; Alsop, Jim et al. (2018) Relebactam Is a Potent Inhibitor of the KPC-2 ?-Lactamase and Restores Imipenem Susceptibility in KPC-Producing Enterobacteriaceae. Antimicrob Agents Chemother 62:
Tsalik, Ephraim L; Bonomo, Robert A; Fowler Jr, Vance G (2018) New Molecular Diagnostic Approaches to Bacterial Infections and Antibacterial Resistance. Annu Rev Med 69:379-394
Cheng, Zishuo; VanPelt, Jamie; Bergstrom, Alexander et al. (2018) A Noncanonical Metal Center Drives the Activity of the Sediminispirochaeta smaragdinae Metallo-?-lactamase SPS-1. Biochemistry 57:5218-5229
VanPelt, Jamie; Shurina, Ben A; Ramelot, Theresa A et al. (2018) 1H, 13C, and 15N backbone resonance assignments for KPC-2, a class A serine-?-lactamase. Biomol NMR Assign :
Papp-Wallace, Krisztina M; Nguyen, Nhu Q; Jacobs, Michael R et al. (2018) Strategic Approaches to Overcome Resistance against Gram-Negative Pathogens Using ?-Lactamase Inhibitors and ?-Lactam Enhancers: Activity of Three Novel Diazabicyclooctanes WCK 5153, Zidebactam (WCK 5107), and WCK 4234. J Med Chem 61:4067-4086
Ju, Lin-Cheng; Cheng, Zishuo; Fast, Walter et al. (2018) The Continuing Challenge of Metallo-?-Lactamase Inhibition: Mechanism Matters. Trends Pharmacol Sci 39:635-647

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