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 Title: Structural and functional relationship of photosynthetic bacterial reaction centers Author(s): Gao, Jiliang Doctoral Committee Chair(s): Wraight, Colin A. Department / Program: Biophysics and Computational Biology Discipline: Biophysics Degree Granting Institution: University of Illinois at Urbana-Champaign Degree: Ph.D. Genre: Dissertation Subject(s): Biology, Molecular Chemistry, Biochemistry Biophysics, General Abstract: The structure of reaction centers (RCs) was modified with various sulfhydryl reagents. It was found that mercurial reagents, especially p-chloromercuribenzenesulfate (pCMBS), inhibit Q$\sb{\rm B}$ function in RCs from Rhodobacter sphaeroides and Rhodobacter capsulatus, but not in Rhodopseudomonas viridis. By comparison of the protein sequences and structures of RCs of three species, it was suggested that Cys$\sp{\rm L108}$ in Rb. sphaeroides (Cys$\sp{\rm L109}$ in Rb. capsulatus) is the best candidate for the mercurial reagents to inhibit Q$\sb{\rm B}$ function. And a mechanism was suggested. By construction of a mutant of Rb. sphaeroides, in which Cys$\sp{\rm L108}$ was changed to serine using site-directed mutagenesis, this suggestion was strongly supported by the absence of any mercurial reagent effect on this mutant.Charge recombination kinetics and thermodynamics equilibrium involving the high potential c-type cytochromes (C$\sb{\rm H1}$ and C$\sb{\rm H2}$) and quinone Q$\sb{\rm A}$ in RCs from Rp. viridis were investigated. It was suggested that the charge recombinations have two pathways, a direct pathway involving long distance electron transfer between Q$\sb{\rm A}\sp-$ and either C$\sb{\rm H1}\sp+$ and C$\sb{\rm H2}\sp+$, and indirect pathway, via thermal repopulation of P$\sp+$ Q$\sb{\rm A}\sp-$ followed by recombination of that state. From the schemes, we calculated the decay rates of C$\rm\sb{H2}\sp+C\sb{H1}Q\sb{A}\sp-$ and C$\rm\sb{H2}\sp+C\sb{H1}\sp+Q\sb{A}\sp-$ charge recombinations. In a wide pH and temperature range, the calculated rates fit the experimental data very well. This result indicates that there are no significant electrostatic interactions between the relevant redox centers in isolated RCs.Heterogeneity was observed in the $\rm P\sp+Q\sb{B}\sp-$ recombination of RCs from Rp. viridis. The kinetics of the $\rm P\sp+Q\sb{B}\sp-$ recombination were well fitted by two exponential components. The two components were interconvertible, and pH, temperature and salt dependent. The presence of two long-lived, kinetically distinct conformations of the RCs was suggested. Calculation of the equilibrium constant of electron transfer between Q$\sb{\rm A}$ and Q$\sb{\rm B}$, K$\sb2$, for both components, showed that the two values were almost the same over a wide range of pH. This result suggests that the source of heterogeneity is in the energetic level of intermediate state $\rm P\sp+I\sp-$ through which both $\rm P\sp+Q\sb{A}\sp-$ and $\rm P\sp+Q\sb{B}\sp-$ recombine. Based on the pH dependences of K$\sb2$ and the conformational equilibrium constant, K$\sb{\rm cs}$, a mechanism of the heterogeneity was proposed. Issue Date: 1991 Type: Text Language: English URI: http://hdl.handle.net/2142/22147 Rights Information: Copyright 1991 Gao, Jiliang Date Available in IDEALS: 2011-05-07 Identifier in Online Catalog: AAI9124416 OCLC Identifier: (UMI)AAI9124416
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