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Lishan Liu PhD dissertation.pdf (4.09 MB)

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DEVELOPMENT AND APPLICATION OF A NOVEL PULSED EPR APPROACH FOR MEMBRANE PROTEIN LOCAL SECONDARY STRUCTURE CHARACTERIZATION

liu, lishan
http://rave.ohiolink.edu/etdc/view?acc_num=miami1442394527

Abstract Details

2015, Doctor of Philosophy, Miami University, Chemistry and Biochemistry.
Membrane proteins perform a variety of important biological functions which are essential to the survival of organisms. However, due to their inherent hydrophobic nature and complexity of their native environments, it is extremely difficult to gain structural information of full-length membrane proteins in their native-like environments using traditional biophysical techniques. This dissertation presents the development and application of a novel local secondary structure approach for membrane proteins and peptides using spin label EPR spectroscopy. Utilizing site-direct spin-labeling (SDSL) and Electron Spin Echo Envelope Modulation (ESEEM), local secondary structure of membrane proteins and peptides can be identified with µg scale of sample within a couple of hours. Background of this methodology can be found in Chapter 1. Detailed experimental procedure and data analysis of this ESEEM approach are described in Chapter 2. In the following chapters, this ESEEM approach was used to map out a model transmembrane a-helical peptide embedded in bicelles (chapter 3, 4 and 5). We demonstrated that a unique ESEEM pattern can be obtained with multiple 2H-labeled amino acids to identify a-helical structural components. Also, it was shown with model peptides that an a-helix and a ß-strand can be distinguished via this ESEEM approach (Chapter 6). Moreover, we showed that this technique can be extended to probe the local secondary structure of full-length proteins that were difficult to study with other biophysical techniques. Two different sites of the full length human vimentin protein with available X-ray structures from fragments was studied with this ESEEM approach and local structural information was obtained (Chapter 7). Final remarks and future directions are also discussed to conclude this study (Chapter 8).
Gary A. Lorigan, Dr. (Advisor)
Carole Dabney-Smith, Dr. (Committee Chair)
C. Scott Hartley, Dr. (Committee Member)
David L. Tierney, Dr. (Committee Member)
Mitchell F. Balish, Dr. (Committee Member)
138 p.
Biochemistry; Biophysics; Chemistry
EPR, ESEEM, Membrane protein structure, Secondary strucuture

Recommended Citations

Citations

  • liu, L. (2015). DEVELOPMENT AND APPLICATION OF A NOVEL PULSED EPR APPROACH FOR MEMBRANE PROTEIN LOCAL SECONDARY STRUCTURE CHARACTERIZATION [Doctoral dissertation, Miami University]. OhioLINK Electronic Theses and Dissertations Center. http://rave.ohiolink.edu/etdc/view?acc_num=miami1442394527

    APA Style (7th edition)

  • liu, lishan. DEVELOPMENT AND APPLICATION OF A NOVEL PULSED EPR APPROACH FOR MEMBRANE PROTEIN LOCAL SECONDARY STRUCTURE CHARACTERIZATION. 2015. Miami University, Doctoral dissertation. OhioLINK Electronic Theses and Dissertations Center, http://rave.ohiolink.edu/etdc/view?acc_num=miami1442394527.

    MLA Style (8th edition)

  • liu, lishan. "DEVELOPMENT AND APPLICATION OF A NOVEL PULSED EPR APPROACH FOR MEMBRANE PROTEIN LOCAL SECONDARY STRUCTURE CHARACTERIZATION." Doctoral dissertation, Miami University, 2015. http://rave.ohiolink.edu/etdc/view?acc_num=miami1442394527

    Chicago Manual of Style (17th edition)

miami1442394527
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© 2015, all rights reserved.
This open access ETD is published by Miami University and OhioLINK.