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Single-Molecule Spectroscopy And Imaging Studies Of Protein Folding-Unfolding Conformational Dynamics: The Multiple-State And Multiple-Channel Energy Landscape

Wang, Zijian
http://rave.ohiolink.edu/etdc/view?acc_num=bgsu1459942296

Abstract Details

2016, Doctor of Philosophy (Ph.D.), Bowling Green State University, Photochemical Sciences.
Protein conformational dynamics often plays a critical role in protein functions. We have characterized the spontaneous folding-unfolding conformational fluctuation dynamics of calmodulin (CaM) at thermodynamic equilibrium conditions by using single-molecule fluorescence resonance energy transfer (FRET) spectroscopy. We studied protein folding dynamics under simulated biological conditions to gain a deep, mechanistic understanding of this important biological process. We have identified multiple folding transition pathways and characterized the underlying energy landscape of the single-molecule protein conformational fluctuation trajectories. Our results suggest that the folding dynamics of CaM molecules involves a complex multiple-pathway multiple-state energy landscape, rather than an energy landscape of two-state dynamical process. Our probing single-molecule FRET fluctuation experiments demonstrate a new approach of studying spontaneous protein folding-unfolding conformational dynamics at the equilibrium that features recording long time single-molecule conformational fluctuation trajectories. This technique yields rich statistical and dynamical information far beyond traditional ensemble-averaged measurements. We characterize the conformational dynamics of single CaM interacting with C28W. The single CaM molecules are partially unfolded by GdmCl, and the folded and unfolded CaM molecules are approximately equally populated. Under this condition, the majority of the single protein CaM undergoes spontaneous folding-unfolding conformational fluctuations. Using single molecule FRET spectroscopy, we study each of the single protein’s conformational dynamics inthe presence of C28W-CaM interactions. The results show an interesting folding-upon-binding dynamic process, and a conformational selection mechanism is further confirmed. The effect of molecular crowding on protein folding process is a key issue in the understanding of protein folding dynamics in living cells. Due to the complexity and interplay between various interactions existing in an equally favored environment of protein folding and unfolding conformational dynamics, such simple reduced entropic enhancement model do not suffice in describing protein folding conformational dynamics. We observe, at higher concentration of crowding reagent Ficoll 70, single protein molecules spontaneously denature into unfolded proteins which involves a combined process of polymer-polymer interaction, entropic effects and solvation thermodynamics and dynamics. Such heterogeneous unfolding process can serve as a first step to a mechanistic understanding of living cell disease as a result of molecular crowding effect, protein aggregates and fibril formation.
H. Peter Lu, Dr. (Advisor)
Gabriela Bidart-Bouzat, Dr. (Committee Member)
Ksenija D. Glusac, Dr. (Committee Member)
George Bullerjahn, Dr. (Committee Member)
132 p.
Chemistry
Protein Folding; Single-Molecule Spectroscopy; Protein-Protein Interactions; Condensed Phase Dynamics

Recommended Citations

Citations

  • Wang, Z. (2016). Single-Molecule Spectroscopy And Imaging Studies Of Protein Folding-Unfolding Conformational Dynamics: The Multiple-State And Multiple-Channel Energy Landscape [Doctoral dissertation, Bowling Green State University]. OhioLINK Electronic Theses and Dissertations Center. http://rave.ohiolink.edu/etdc/view?acc_num=bgsu1459942296

    APA Style (7th edition)

  • Wang, Zijian. Single-Molecule Spectroscopy And Imaging Studies Of Protein Folding-Unfolding Conformational Dynamics: The Multiple-State And Multiple-Channel Energy Landscape. 2016. Bowling Green State University, Doctoral dissertation. OhioLINK Electronic Theses and Dissertations Center, http://rave.ohiolink.edu/etdc/view?acc_num=bgsu1459942296.

    MLA Style (8th edition)

  • Wang, Zijian. "Single-Molecule Spectroscopy And Imaging Studies Of Protein Folding-Unfolding Conformational Dynamics: The Multiple-State And Multiple-Channel Energy Landscape." Doctoral dissertation, Bowling Green State University, 2016. http://rave.ohiolink.edu/etdc/view?acc_num=bgsu1459942296

    Chicago Manual of Style (17th edition)

bgsu1459942296
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© 2016, some rights reserved.Creative Commons License Single-Molecule Spectroscopy And Imaging Studies Of Protein Folding-Unfolding Conformational Dynamics: The Multiple-State And Multiple-Channel Energy Landscape by Zijian Wang is licensed under a Creative Commons Attribution-NonCommercial-ShareAlike 3.0 Unported License. Based on a work at etd.ohiolink.edu.
This open access ETD is published by Bowling Green State University and OhioLINK.