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PHD Thesis Yangzhong Qin.pdf (8 MB)
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Abstract Header
Ultrafast Hydration Dynamics Probed by Tryptophan at Protein Surface and Protein-DNA Interface
Author Info
Qin, Yangzhong
Permalink:
http://rave.ohiolink.edu/etdc/view?acc_num=osu1416998263
Abstract Details
Year and Degree
2015, Doctor of Philosophy, Ohio State University, Physics.
Abstract
As we all live in a special water planet Earth, the significance of water to life has been universally recognized. The reason why water is so important to life has intrigued many researchers. This dissertation will focus on the ultrafast dynamics of protein surface water and protein-DNA interfacial water which have direct importance to the protein structure and function. Using tryptophan as an intrinsic fluorescence probe, combined with site-directed mutagenesis and ultrafast fluorescence up-conversion spectroscopy, we can achieve single residue spatial resolution and femtosecond temporal resolution. We can also precisely determine the local hydration water dynamics by monitoring the Stokes shift of tryptophan one at a time. Previously, the protein surface hydration has been extensively studied by our group. In this thesis, we will provide more details on the methods we are using to extract the hydration dynamics, and also validate our methods from both experimental and theoretical perspectives. To further interrogate the interfacial water hydration dynamics relative to the protein surface hydration, we studied two DNA polymerases: DNA Polymerase IV (Dpo4) and DNA Polymerase Beta (Pol ß). Both proteins show typical surface hydration pattern with three distinct time components including: (i) the ultrafast sub-picosecond component reflects the bulk type water motion; (ii) a few picoseconds component shows the inner water relaxation mainly corresponding to the local libration and reorientation; (iii) the tens to hundred picoseconds component represents the water-protein coupled motion involving the whole water network reorganization. Dpo4, a loosely DNA binding protein, exhibits very flexible interfacial water which resembles its surface water yet with a significantly reduced ultrafast component. Such dynamic interfacial water not only maintains interfacial flexibility, but also contributes to the low fidelity of the protein. In contrast to the Dpo4, pol ß tightly binds to the DNA with higher fidelity. The interfacial water is about 2 times slower than the surface water. Most importantly, the ultrafast component disappeared for the interfacial water indicating a strongly confined local environment. To further inspect the surface and interfacial water property, we carried out a 4 ns MD simulation, which reveals that all the interfacial water forms only one to two hydration layers from the protein and DNA surface, and no bulk type water was discovered. For both enzymes, the interfacial water is still fluctuating on picoseconds time scales to facilitate their dynamic function including substrate binding, protein sliding on DNA and dNTP sampling. Despite many hydration dynamics were studied at room temperature, some enzymes are optimally functioning at much higher temperature. Dpo4 was originally found on Sulfolobus solfataricus, which grows in volcanic hot springs with temperature around 75-90 °C. This inspired us to study its hydration temperature dependence from 1 °C to 60 °C. Simple Arrhenius equation can fit all the data for each mutant and determine the activation enthalpies ranging from 5 kJ/mol to 15 kJ/mol. We also found a clear correlation between the solvation and tryptophan side chain motion, strongly supporting the coupled motion between water and protein and the famous slaving model.
Committee
Dongping Zhong (Advisor)
David Stroud (Committee Member)
Fengyuan Yang (Committee Member)
Jay Gupta (Committee Member)
Pages
152 p.
Subject Headings
Biochemistry
;
Biophysics
;
Physics
Keywords
Ultrafast Spectroscopy, Solvation Dynamics, Hydration Dynamics, Water Motions, Interfacial Water, Temperature Dependence, Activation Energy, Protein Side Chain Motion
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Citations
Qin, Y. (2015).
Ultrafast Hydration Dynamics Probed by Tryptophan at Protein Surface and Protein-DNA Interface
[Doctoral dissertation, Ohio State University]. OhioLINK Electronic Theses and Dissertations Center. http://rave.ohiolink.edu/etdc/view?acc_num=osu1416998263
APA Style (7th edition)
Qin, Yangzhong.
Ultrafast Hydration Dynamics Probed by Tryptophan at Protein Surface and Protein-DNA Interface.
2015. Ohio State University, Doctoral dissertation.
OhioLINK Electronic Theses and Dissertations Center
, http://rave.ohiolink.edu/etdc/view?acc_num=osu1416998263.
MLA Style (8th edition)
Qin, Yangzhong. "Ultrafast Hydration Dynamics Probed by Tryptophan at Protein Surface and Protein-DNA Interface." Doctoral dissertation, Ohio State University, 2015. http://rave.ohiolink.edu/etdc/view?acc_num=osu1416998263
Chicago Manual of Style (17th edition)
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Document number:
osu1416998263
Download Count:
644
Copyright Info
© 2015, all rights reserved.
This open access ETD is published by The Ohio State University and OhioLINK.