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Biochemical Characterization of Glutathione Transferase YliJ from Escherichia coli

Aboagye, Collins
http://rave.ohiolink.edu/etdc/view?acc_num=ysu1453197715

Abstract Details

2015, Master of Science in Chemistry, Youngstown State University, Department of Chemistry.
Glutathione transferases, or GSTs, protect cellular components from the dangers posed by electrophiles by catalyzing the conjugation of the electrophilic molecules with glutathione. This makes GSTs very successful detoxifying agents in most organisms. Glutathione transferase YliJ, also known as GstB, has been postulated to catalyze the glutathione mediated dehalogenation of bromoacetate. The objective of this research is to develop a simple purification procedure for YliJ, design a safe method for assaying YliJ activity, characterize YliJ, and to search for other potential substrates for YliJ. YliJ was overexpressed in E. coli cells and purified using ammonium sulfate precipitation and ion exchange chromatography. The presence and purity of YliJ was verified by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. YliJ has a KM of 1.60 mM, a kcat of 175.0 s-1 and a catalytic efficiency of 1.09 x 105 M-1s-1 for GSH when conjugated to bromoacetate. In comparison, YliJ has a KM of 0.1753 mM, a kcat of 155.3 s-1, and a catalytic efficiency of 8.86 x 104 M-1s-1 for GSH when conjugated to iodoacetate. E. coli cells expressing YliJ protein and the knockout strain BW25113ΔyliJ lacking YliJ were tested for sensitivity to halogenated compounds and antibiotics. The results strongly support the suggestion that a small molecule containing a carboxylate moiety could be the physiological substrate of YliJ.
Nina Stourman, PhD (Advisor)
Michael Serra, PhD (Committee Member)
Josef Simeonsson, PhD (Committee Member)
75 p.
Biochemistry; Chemistry; Microbiology
Chemistry; Biochemistry; Glutathione transferase; Escherichia coli

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Citations

  • Aboagye, C. (2015). Biochemical Characterization of Glutathione Transferase YliJ from Escherichia coli [Master's thesis, Youngstown State University]. OhioLINK Electronic Theses and Dissertations Center. http://rave.ohiolink.edu/etdc/view?acc_num=ysu1453197715

    APA Style (7th edition)

  • Aboagye, Collins. Biochemical Characterization of Glutathione Transferase YliJ from Escherichia coli. 2015. Youngstown State University, Master's thesis. OhioLINK Electronic Theses and Dissertations Center, http://rave.ohiolink.edu/etdc/view?acc_num=ysu1453197715.

    MLA Style (8th edition)

  • Aboagye, Collins. "Biochemical Characterization of Glutathione Transferase YliJ from Escherichia coli." Master's thesis, Youngstown State University, 2015. http://rave.ohiolink.edu/etdc/view?acc_num=ysu1453197715

    Chicago Manual of Style (17th edition)

ysu1453197715
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© 2015, all rights reserved.
This open access ETD is published by Youngstown State University and OhioLINK.