Student Summer Scholars
 

Keywords

bacillithiol, BshC, fluorescense assays, mutagenesis, fosfomycin resistance, crystallography

Disciplines

Chemistry

Included in

Chemistry Commons

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Abstract

BshC is the final enzyme in a three step pathway for the synthesis of bacillithiol, a compound that enables resistance to fosfomycin in Gram-positive bacteria. BshC is unique from other enzymes of its kind because of an additional ADP binding site and inactivity when studied in the laboratory. To explore BshC function, several site-directed mutants have been selected within the ADP binding pocket. Fluorescence assays have been utilized on the wild-type BshC and one mutant, Y510Q. We determined that Y510Q does not bind ATP as effectively as wild-type BshC. These fluorescence assays will be utilized on W506L, E384A, and H386A mutants and structural analysis of all the mutants will be initiated. Gaining more understanding of the structure of these mutants and how they bind ATP will give a better understanding of how BshC binds its substrate, which will allow the development of inhibitors to combat fosfomycin resistance.