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Characterization of L1, the metallo-Β-lactamase from Stenotrophomonas maltophilia

Garrity, James D.
http://rave.ohiolink.edu/etdc/view?acc_num=miami1092150871

Abstract Details

2004, Doctor of Philosophy, Miami University, Chemistry and Biochemistry.
Zinc-containing metallo-Β-lactamases are an emerging class of enzymes that render bacteria resistant to Β-lactam-containing antibiotics. In an effort to better understand the function of the metallo-Β-lactamase L1 from Stenotrophomonas maltoplia, spectroscopic and mechanistic studies were performed. Mutagenesis and biochemical assays of nine L1 active site residues, predicted from computational studies to be important for tight substrate binding to the enzyme, revealed that none of the residues play a significant role in the binding of substrate to L1. Investigation of a metal-binding aspartic acid showed that, in addition to ligating a Zn(II), this residue also plays an important role in catalytic cycle of the enzyme by properly orienting a water molecule that takes part in a rate-limiting protonation event. Fluorescence studies revealed that the rate of motion of a flexible loop of amino acids that extends over the active site of L1 correlates to the formation of a non-rate-limiting intermediate in the catalytic cycle of L1. Rapid-scanning and rapid-freeze quench EPR spectrophotometry studies provided the first direct evidence that the reaction intermediate of L1 is metal bound. This dissertation offers data, which, along with previous results on L1 and other metallo-Β-lactamases, can be integrated and used to guide further rational inhibitor design efforts.
Michael Crowder (Advisor)
198 p.
Chemistry, General
L1; Wild Type L1; nitrocefin; kcat; METALLO-¿¿¿¿-LACTAMASE; Mutants

Recommended Citations

Citations

  • Garrity, J. D. (2004). Characterization of L1, the metallo-Β-lactamase from Stenotrophomonas maltophilia [Doctoral dissertation, Miami University]. OhioLINK Electronic Theses and Dissertations Center. http://rave.ohiolink.edu/etdc/view?acc_num=miami1092150871

    APA Style (7th edition)

  • Garrity, James. Characterization of L1, the metallo-Β-lactamase from Stenotrophomonas maltophilia. 2004. Miami University, Doctoral dissertation. OhioLINK Electronic Theses and Dissertations Center, http://rave.ohiolink.edu/etdc/view?acc_num=miami1092150871.

    MLA Style (8th edition)

  • Garrity, James. "Characterization of L1, the metallo-Β-lactamase from Stenotrophomonas maltophilia." Doctoral dissertation, Miami University, 2004. http://rave.ohiolink.edu/etdc/view?acc_num=miami1092150871

    Chicago Manual of Style (17th edition)

miami1092150871
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© 2004, all rights reserved.
This open access ETD is published by Miami University and OhioLINK.