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Identification of Potential TonB-Interactive Sites in the Periplasmic Domain of the ExbD Protein

Cholewa, Kelly M.
http://rave.ohiolink.edu/etdc/view?acc_num=bgsu1479398493813069

Abstract Details

2016, Master of Science (MS), Bowling Green State University, Biological Sciences.
The TonB energy transduction complex (TonB, ExbB, and ExbD) couples the proton motive force (PMF) to acquire and transport iron in gram-negative bacteria. ExbD is needed to gather energy from the PMF and facilitates conformational changes in the TonB C-terminus. The goal of this study was to determine possible interaction sites between ExbD and TonB. Site-directed mutagenesis was performed on possible interaction sites to create three constructs that selected specific E. coli exbD codons and were replaced with Y. enterocolitica exbD codons. The codons were chosen by identifying residues that diverged significantly in the ExbD protein compared to homologues in the family Enterobacteriaceae. Structural predictions were generated to compare the periplasmic space of ExbD of the homologues. The solvent accessibility for E. coli ExbD was generated from the structural predictions with surface residues as possible contact sites. The function of each construct was assessed by a spot titer assay of sensitivity to four group B colicins: M, B, Ia, and D. My results showed that two of these codon changes did not significantly affect the ability of ExbD to support TonB-dependent uptake of colicins, while a third codon change reduced sensitivity to one of the four colicins tested.
Raymond Larsen, Ph.D (Advisor)
George Bullerjahn, Ph.D (Committee Member)
Vipaporn Phuntumart, Ph.D (Committee Member)
77 p.
Biology; Microbiology; Molecular Biology
ExbD; TonB System; Site-Directed Mutagenesis; E coli

Recommended Citations

Citations

  • Cholewa, K. M. (2016). Identification of Potential TonB-Interactive Sites in the Periplasmic Domain of the ExbD Protein [Master's thesis, Bowling Green State University]. OhioLINK Electronic Theses and Dissertations Center. http://rave.ohiolink.edu/etdc/view?acc_num=bgsu1479398493813069

    APA Style (7th edition)

  • Cholewa, Kelly. Identification of Potential TonB-Interactive Sites in the Periplasmic Domain of the ExbD Protein. 2016. Bowling Green State University, Master's thesis. OhioLINK Electronic Theses and Dissertations Center, http://rave.ohiolink.edu/etdc/view?acc_num=bgsu1479398493813069.

    MLA Style (8th edition)

  • Cholewa, Kelly. "Identification of Potential TonB-Interactive Sites in the Periplasmic Domain of the ExbD Protein." Master's thesis, Bowling Green State University, 2016. http://rave.ohiolink.edu/etdc/view?acc_num=bgsu1479398493813069

    Chicago Manual of Style (17th edition)

bgsu1479398493813069
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© 2016, all rights reserved.
This open access ETD is published by Bowling Green State University and OhioLINK.