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Challenges in Characterizing Membrane Proteins and Intrinsically Disordered Regions Involved in Mycobacterium Tuberculosis Cell Division

Title: Challenges in Characterizing Membrane Proteins and Intrinsically Disordered Regions Involved in Mycobacterium Tuberculosis Cell Division.
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Name(s): Escobar Bravo, Cristian Andres, author
Cross, Timothy A., professor directing dissertation
Jones, Kathryn M. (Kathryn Marjorie), university representative
Fajer, Peter G., committee member
Logan, Timothy M., 1961-, committee member
Mattoussi, Hedi, committee member
Stagg, Scott M., committee member
Florida State University, degree granting institution
College of Arts and Sciences, degree granting college
Institute of Molecular Biophysics , degree granting department
Type of Resource: text
Genre: Text
Doctoral Thesis
Issuance: monographic
Date Issued: 2017
Publisher: Florida State University
Place of Publication: Tallahassee, Florida
Physical Form: computer
online resource
Extent: 1 online resource (222 pages)
Language(s): English
Abstract/Description: Structural biology has been successful in the characterization of soluble protein, which has given insight in the inner working of several thousands of proteins. However, structural characterization of proteins has been limited in two areas. One area corresponds to the rise on interest on intrinsically disorder proteins. This kind of proteins has changed the structure-function paradigm since they can perform important cellular function without a defined three dimensional structure. The second are is the characterization of membrane proteins, which contrary to soluble proteins, they are embedded in an anisotropic environment that makes them difficult to characterize. This dissertation work was focused in the characterization of two important membrane proteins involved in cell division of Mycobacterium tuberculosis, ChiZ and FtsX. The first part of this work is related to the characterization of ChiZ, a cell division protein possibly involved in peptidoglycan remodeling and control of cell division progression. Structural characterization of ChiZ was mainly focused on the N-terminal soluble region that is intrinsically disordered. This region was previously identified to have peptidoglycan hydrolysis activity. Thus, a combination of solution and solid state NMR to characterize the structure of ChiZ N-terminal region in solution and in the full length protein reconstituted in liposomes. In addition to solution NMR, ChiZ N-terminal intrinsically disordered region was characterized with other method including circular dichroism, dynamic light scattering, size exclusion chromatography and small angle X-ray scattering. Thus, the objective was to obtain a clear picture of the behavior of the intrinsically disordered region in solution. Furthermore, characterization of ChiZ peptidoglycan hydrolysis activity was attempted with the goal of correlating structural properties of the N-terminal region and the protein function. The second part of this study focuses on the challenges of working with membrane proteins. The subject of study was FtsX, a membrane protein is one of the components of the cell division machinery or divisome. This part of the work attempts to give a general guideline for optimizing membrane protein purification and reconstitution with large emphasis on detergent selection. Detergent selection can affect each step of FtsX work, from protein purification to membrane protein reconstitution and sample preparation for solid state NMR experiments.
Identifier: FSU_2017SP_EscobarBravo_fsu_0071E_13765 (IID)
Submitted Note: A Dissertation submitted to the Institute of Molecular Biophysics in partial fulfillment of the Doctor of Philosophy.
Degree Awarded: Spring Semester 2017.
Date of Defense: March 23, 2017.
Keywords: Intrinsically disordered proteins, Membrane proteins, Nuclear magnetic resonance
Bibliography Note: Includes bibliographical references.
Advisory Committee: Timothy A. Cross, Professor Directing Dissertation; Kathryn M. Jones, University Representative; Peter Fajer, Committee Member; Timothy Logan, Committee Member; Hedi Mattoussi, Committee Member; Scott Stagg, Committee Member.
Subject(s): Biophysics
Biochemistry
Persistent Link to This Record: http://purl.flvc.org/fsu/fd/FSU_2017SP_EscobarBravo_fsu_0071E_13765
Owner Institution: FSU

Choose the citation style.
Escobar Bravo, C. A. (2017). Challenges in Characterizing Membrane Proteins and Intrinsically Disordered Regions Involved in Mycobacterium Tuberculosis Cell Division. Retrieved from http://purl.flvc.org/fsu/fd/FSU_2017SP_EscobarBravo_fsu_0071E_13765