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MS thesis- Rehani Perera.pdf (80.42 MB)

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Determining the Structural Dynamics and Topology of Canonical HOLIN-S05 Using EPR Spectroscopy

Perera, Rehani Shinuka
http://orcid.org/0000-0001-8312-5586
http://rave.ohiolink.edu/etdc/view?acc_num=miami1591797430542798

Abstract Details

2020, Master of Science, Miami University, Chemistry and Biochemistry.
Holins are membrane proteins encoded by bacteriophages for promoting cell lysis by forming fatal "holes." In phage infections of gram-negative bacteria cell lysis occurs destroying the three layers of the cell envelope: cytoplasmic membrane, peptidoglycan layer, and outer membrane by Holin, Endolysin, and Spanin proteins respectively. Holins are the key determinant of the host lysis timing in phage infections. Canonical holins of phage lambda are of two types, actual lethal holin - S105, and antiholin - S107 as a result of the dual start motif of the Sλ gene that encodes them. Previously, the structural dynamics and topology of this system have not been studied using EPR spectroscopy in native membrane mimetics. In this study, SDSL-EPR spectroscopy is used to unravel structural dynamics and topology of the S105 in a micelle, liposome, and SMALPs environments. CW-EPR line shapes of selected MTSL labeled cysteine mutants across the three transmembrane domains and C-terminus of S105 were used to probe the dynamics of S105. CW-EPR power saturation experiments were used to determine the topology of different segments of S105 in a membrane utilizing the accessibility of paramagnetic relaxants O2 and NiEDDA. Successful insertion of S105 in membrane mimetics was confirmed by this study.
Gary Lorigan, Ph.D. (Advisor)
Carole Dabney-Smith, Ph.D. (Committee Chair)
Richard Page, Ph.D. (Committee Member)
Scott Hartley, Ph.D. (Committee Member)
57 p.
Biochemistry; Chemistry; Physical Chemistry
EPR; S105; Canonical Holin; Membrane protein; Protein topology; Protein dynamics;

Recommended Citations

Citations

  • Perera, R. S. (2020). Determining the Structural Dynamics and Topology of Canonical HOLIN-S05 Using EPR Spectroscopy [Master's thesis, Miami University]. OhioLINK Electronic Theses and Dissertations Center. http://rave.ohiolink.edu/etdc/view?acc_num=miami1591797430542798

    APA Style (7th edition)

  • Perera, Rehani. Determining the Structural Dynamics and Topology of Canonical HOLIN-S05 Using EPR Spectroscopy. 2020. Miami University, Master's thesis. OhioLINK Electronic Theses and Dissertations Center, http://rave.ohiolink.edu/etdc/view?acc_num=miami1591797430542798.

    MLA Style (8th edition)

  • Perera, Rehani. "Determining the Structural Dynamics and Topology of Canonical HOLIN-S05 Using EPR Spectroscopy." Master's thesis, Miami University, 2020. http://rave.ohiolink.edu/etdc/view?acc_num=miami1591797430542798

    Chicago Manual of Style (17th edition)

miami1591797430542798
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© 2020, all rights reserved.
This open access ETD is published by Miami University and OhioLINK.