C A Wraight

University of Illinois, Urbana-Champaign, Urbana, Illinois, United States

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Publications (87)320.17 Total impact

  • [Show abstract] [Hide abstract]
    ABSTRACT: Recent studies have shown that only quinones with a 2-methoxy group can act simultaneously as the primary (QA) and secondary (QB) electron acceptors in photosynthetic reaction centers from purple bacteria such as Rb. sphaeroides. (13)C HYSCORE measurements of the 2-methoxy group in the semiquinone states, SQA and SQB, were compared with DFT calculations of the (13)C hyperfine couplings as a function of the 2-methoxy dihedral angle. X-ray structure comparisons support 2-methoxy dihedral angle assignments corresponding to a redox potential gap (ΔE m) between QA and QB of 175-193 mV. A model having a methyl group substituted for the 2-methoxy group exhibits no electron affinity difference. This is consistent with the failure of a 2-methyl ubiquinone analogue to function as QB in mutant reaction centers with a ΔE m of ∼160-195 mV. The conclusion reached is that the 2-methoxy group is the principal determinant of electron transfer from QA to QB in type II photosynthetic reaction centers with ubiquinone serving as both acceptor quinones.
    Journal of Physical Chemistry Letters 08/2014; 5(15):2506-2509. · 6.59 Impact Factor
  • The journal of physical chemistry. B. 07/2014;
  • Melvin Okamura, Colin A. Wraight, Rienk van Grondelle
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    ABSTRACT: This Special Issue of Photosynthesis Research honors Louis M. N. Duysens, Roderick K. Clayton, and George Feher, three pioneering researchers whose work on bacterial photosynthesis laid much of the groundwork for our understanding of the role of the reaction center in photosynthetic light energy conversion. Their key discoveries are briefly summarized and an overview of the special issue is presented.
    Photosynthesis Research 05/2014; 120(1-2). · 3.15 Impact Factor
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    ABSTRACT: The secondary quinone anion radical QB- (SQB) in reaction centers of Rhodobacter sphaeroides interacts with Nδ of His-L190 and Np (peptide nitrogen) of Gly-L225 involved in hydrogen bonds to the QB carbonyls. In this work, S-band (~3.6 GHz) ESEEM was used with the aim of obtaining a complete characterization of the nuclear quadrupole interaction (nqi) tensors for both nitrogens by approaching the cancellation condition between the isotropic hyperfine coupling and 14N Zeeman frequency at lower microwave frequencies. By performing measurements at S-band we found a dominating contribution of Nδ in the form of a zero-field nqi triplet at 0.55 MHz, 0.92 MHz, and 1.47 MHz, defining the quadrupole coupling constant K = e2qQ/4h = 0.4 MHz and associated asymmetry parameter η = 0.69. Estimates of the hyperfine interaction (hfi) tensors for Nδ and Np were obtained from simulations of 1D and 2D 14,15N X-band and three-pulse 14N S-band spectra with all nuclear tensors defined in the SQB g-tensor coordinate system. From simulations we conclude that the contribution of Np to the S-band spectrum is suppressed by its strong nqi and weak isotropic hfi comparable to the level of hyperfine anisotropy, despite the near-cancellation condition for Np at S-band. The excellent agreement between our EPR simulations and DFT calculations of the nitrogen hfi and nqi tensors to SQB is promising for the future application of powder ESEEM to full tensor characterizations.
    The Journal of Physical Chemistry B 01/2014; · 3.61 Impact Factor
  • Source
    Colin A Wraight
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    ABSTRACT: Roderick K. Clayton passed away on October 23, 2011, at the age of 89, shortly after the plan for this dedicatory issue of Photosynthesis Research had been hatched. I had just written a lengthy letter to him to re-establish contact after a hiatus of 2 or 3 years, and to suggest that I visit him to talk about his life. It isn't clear whether he saw the letter or not, but it was found at his home in Santa Rosa, California. Fortunately, Rod has written two memoirs for Photosynthesis Research that not only cover much of his research on reaction centers (Photosynth Res 73:63-71, 2002) but also provide a humorous and honest look at his personal life (Photosynth Res 19:207-224, 1988). I cannot hope to improve on these and will try, instead, to fill in some of the gaps that Rod's own writing has left, and offer some of my own personal recollections over the more recent years.
    Photosynthesis Research 11/2013; · 3.15 Impact Factor
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    ABSTRACT: Only quinones with a 2-methoxy group can act simultaneously as the primary (QA) and secondary (QB) electron acceptors in photosynthetic reaction centers from Rb. sphaeroides. 13C HYSCORE measurements of the 2-methoxy in the semiquinone states, SQA and SQB, were compared with QM calculations of the 13C couplings as a function of dihedral angle. X-ray structures support dihedral angle assignments corre-sponding to a redox potential gap (∆Em) between QA and QB of ~180 mV. This is consistent with the failure of a ubiquinone analog lacking the 2-methoxy to func-tion as QB in mutant reaction centers with a ∆Em ≈ 160-195 mV.
    Biochemistry 09/2013; · 3.38 Impact Factor
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    ABSTRACT: Surface-enhanced IR absorption spectroscopy (SEIRAS) in the ATR configuration has been performed on reaction centers (RCs) from R. sphaeroides. Surface-enhancement is achieved by a thin, structured gold film present on the surface of an ATR crystal. Purified RCs are immobilized as a monolayer on top of the gold film via a poly his-tag engineered to the C-terminal end of the M subunit. Subsequently, the RCs are reconstituted into a lipid bilayer by in situ dialysis. Light-minus-dark absorbance spectra were recorded under continuous illumination using the spectrum in the dark as the reference. A number of strong bands have been observed indicating the excitation of the special pair as well as alterations of quinone/quinol species. Spectra were recorded at different time intervals with and without liposoluble Q10 coreconstituted into the lipid phase. A steady (photostationary) state was approached slowly and bands were found to increase or decrease reversibly on illumination and relaxation. Tentative assignments were made for some bands, based on previous FTIR measurements. The long time scale of these processes was tentatively explained in terms of interprotein reactions of RC molecules.
    The Journal of Physical Chemistry C 07/2013; 117(32):16357. · 4.84 Impact Factor
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    ABSTRACT: Ubiquinone is an almost universal, membrane-associated redox mediator. Its ability to accept either one or two electrons allows it to function in critical roles in biological electron transport. The redox properties of ubiquinone in vivo are determined by its environment in the binding sites of proteins and by the dihedral angle of each methoxy group relative to the ring plane. This is an attribute unique to ubiquinone among natural quinones and could account for its widespread function with many different redox complexes. In this work, we use the photosynthetic reaction center as a model system for understanding the role of methoxy conformations in determining the redox potential of the ubiquinone/semiquinone couple. Despite the abundance of X-ray crystal structures for the reaction center, quinone site resolution has thus far been too low to provide a reliable measure of the methoxy dihedral angles of the primary and secondary quinones, QA and QB. We performed 2D ESEEM (HYSCORE) on isolated reaction centers with ubiquinones (13)C-labeled at the headgroup methyl and methoxy substituents, and have measured the (13)C isotropic and anisotropic components of the hyperfine tensors. Hyperfine couplings were compared to those derived by DFT calculations as a function of methoxy torsional angle allowing estimation of the methoxy dihedral angles for the semiquinones in the QA and QB sites. Based on this analysis, the orientation of the 2-methoxy groups are distinct in the two sites, with QB more out of plane by 20-30°. This corresponds to an ≈50 meV larger electron affinity for the QB quinone, indicating a substantial contribution to the experimental difference in redox potentials (60-75 mV) of the two quinones. The methods developed here can be readily extended to ubiquinone-binding sites in other protein complexes.
    Biochemistry 06/2013; · 3.38 Impact Factor
  • Biophysical Journal; 01/2013
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    ABSTRACT: In the QB site of the Rba. sphaeroides photosynthetic reaction centre the donation of a hydrogen bond from the hydroxyl group of Ser-L223 to the ubisemiquinone formed after the first flash is debatable. In this study we use a combination of spectroscopy and quantum mechanics/molecular mechanics (QM/MM) calculations to comprehensively explore this topic. We show that ENDOR, ESEEM and HYSCORE spectroscopic differences between the mutant L223SA and the wild type sample (WT) are negligible, indicating only minor perturbations in the ubisemiquinone spin density for the mutant sample. Qualitatively this suggests that a strong hydrogen bond does not exist in the WT between the Ser-L223 hydroxyl group and the semiquinone O1 atom, as removal of this hydrogen bond in the mutant should cause a significant redistribution of spin density in the semiquinone. We show quantitatively, using QM/MM calculations, that a WT model in which the Ser-L223 hydroxyl group is rotated to prevent hydrogen bond formation with the O1 atom of the semiquinone predicts negligible change for the L223SA mutant. This, together with the better agreement between key QM/MM calculated and experimental hyperfine couplings for the non-hydrogen bonded model, leads us to conclude that no strong hydrogen bond is formed between the Ser-L223 hydroxyl group and the semiquinone O1 atom after the first flash. The implications of this finding for quinone reduction in photosynthetic reaction centres are discussed.
    Biochemistry 09/2012; · 3.38 Impact Factor
  • Aidas J. Mattis, Colin A. Wraight
    Biophysical Journal 01/2012; 102(3):575-. · 3.67 Impact Factor
  • Biophysical Journal 01/2012; 102(3):466-. · 3.67 Impact Factor
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    ABSTRACT: In the photosynthetic reaction center from Rhodobacter sphaeroides, the primary (Q(A)) and secondary (Q(B)) electron acceptors are both ubiquinone-10, but with very different properties and functions. To investigate the protein environment that imparts these functional differences, we have applied X-band HYSCORE, a 2D pulsed EPR technique, to characterize the exchangeable protons around the semiquinone (SQ) in the Q(A) and Q(B) sites, using samples of (15)N-labeled reaction centers, with the native high spin Fe(2+) exchanged for diamagnetic Zn(2+), prepared in (1)H(2)O and (2)H(2)O solvent. The powder HYSCORE method is first validated against the orientation-selected Q-band ENDOR study of the Q(A) SQ by Flores et al. (Biophys. J.2007, 92, 671-682), with good agreement for two exchangeable protons with anisotropic hyperfine tensor components, T, both in the range 4.6-5.4 MHz. HYSCORE was then applied to the Q(B) SQ where we found proton lines corresponding to T ≈ 5.2, 3.7 MHz and T ≈ 1.9 MHz. Density functional-based quantum mechanics/molecular mechanics (QM/MM) calculations, employing a model of the Q(B) site, were used to assign the observed couplings to specific hydrogen bonding interactions with the Q(B) SQ. These calculations allow us to assign the T = 5.2 MHz proton to the His-L190 N(δ)H···O(4) (carbonyl) hydrogen bonding interaction. The T = 3.7 MHz spectral feature most likely results from hydrogen bonding interactions of O1 (carbonyl) with both Gly-L225 peptide NH and Ser-L223 hydroxyl OH, which possess calculated couplings very close to this value. The smaller 1.9 MHz coupling is assigned to a weakly bound peptide NH proton of Ile-L224. The calculations performed with this structural model of the Q(B) site show less asymmetric distribution of unpaired spin density over the SQ than seen for the Q(A) site, consistent with available experimental data for (13)C and (17)O carbonyl hyperfine couplings. The implications of these interactions for Q(B) function and comparisons with the Q(A) site are discussed.
    Journal of the American Chemical Society 03/2011; 133(14):5525-37. · 10.68 Impact Factor
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    ABSTRACT: Naturally occurring photosynthetic systems use elaborate pathways of self-repair to limit the impact of photo-damage. Here, we demonstrate a complex consisting of two recombinant proteins, phospholipids and a carbon nanotube that mimics this process. The components self-assemble into a configuration in which an array of lipid bilayers aggregate on the surface of the carbon nanotube, creating a platform for the attachment of light-converting proteins. The system can disassemble upon the addition of a surfactant and reassemble upon its removal over an indefinite number of cycles. The assembly is thermodynamically metastable and can only transition reversibly if the rate of surfactant removal exceeds a threshold value. Only in the assembled state do the complexes exhibit photoelectrochemical activity. We demonstrate a regeneration cycle that uses surfactant to switch between assembled and disassembled states, resulting in an increased photoconversion efficiency of more than 300% over 168 hours and an indefinite extension of the system lifetime.
    Nature Chemistry 11/2010; 2(11):929-36. · 21.76 Impact Factor
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    ABSTRACT: The interaction of cytochrome c with ubiquinol-cytochrome c oxidoreductase (bc₁ complex) has been studied for >30 years, yet many aspects remain unclear or controversial. We report the first molecular dynamic simulations of the cyt c-bc₁ complex interaction. Contrary to the results of crystallographic studies, our results show that there are multiple dynamic hydrogen bonds and salt bridges in the cyt c-c₁ interface. These include most of the basic cyt c residues previously implicated in chemical modification studies. We suggest that the static nature of x-ray structures can obscure the quantitative significance of electrostatic interactions between highly mobile residues. This provides a clear resolution of the discrepancy between the structural data and functional studies. It also suggests a general need to consider dynamic interactions of charged residues in protein-protein interfaces. In addition, a novel structural change in cyt c is reported, involving residues 21-25, which may be responsible for cyt c destabilization upon binding. We also propose a mechanism of interaction between cyt c₁ monomers responsible for limiting the binding of cyt c to only one molecule per bc₁ dimer by altering the affinity of the cytochrome c binding site on the second cyt c₁ monomer.
    Biophysical Journal 10/2010; 99(8):2647-56. · 3.67 Impact Factor
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    ABSTRACT: Photosynthetic reaction centers from Rhodobacter sphaeroides have identical ubiquinone-10 molecules functioning as primary (Q(A)) and secondary (Q(B)) electron acceptors. X-band 2D pulsed EPR spectroscopy, called HYSCORE, was applied to study the interaction of the Q(B) site semiquinone with nitrogens from the local protein environment in natural and (15)N uniformly labeled reactions centers. (14)N and (15)N HYSCORE spectra of the Q(B) semiquinone show the interaction with two nitrogens carrying transferred unpaired spin density. Quadrupole coupling constants estimated from (14)N HYSCORE spectra indicate them to be a protonated nitrogen of an imidazole residue and amide nitrogen of a peptide group. (15)N HYSCORE spectra allowed estimation of the isotropic and anisotropic couplings with these nitrogens. From these data, we calculated the unpaired spin density transferred onto 2s and 2p orbitals of nitrogen and analyzed the contribution of different factors to the anisotropic hyperfine tensors. The hyperfine coupling of other protein nitrogens with the semiquinone is weak (<0.1 MHz). These results clearly indicate that the Q(B) semiquinone forms hydrogen bonds with two nitrogens and provide quantitative characteristics of the hyperfine couplings with these nitrogens, which can be used in theoretical modeling of the Q(B) site. On the basis of the quadrupole coupling constant, one nitrogen can only be assigned to N(delta) of His-L190, consistent with all existing structures. However, we cannot specify between two candidates the residue corresponding to the second nitrogen. Further work employing multifrequency spectroscopic approaches or selective isotope labeling would be desirable for unambiguous assignment of this nitrogen.
    Journal of the American Chemical Society 08/2010; 132(33):11671-7. · 10.68 Impact Factor
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    ABSTRACT: The kinetics of imidazole (Im) and N-methylimidazole (MeIm) binding to oxidized cytochrome (cyt) c(1) of detergent-solubilized bc(1) complex from Rhodobacter sphaeroides are described. The rate of formation of the cyt c(1)-Im complex exhibited three separated regions of dependence on the concentration of imidazole: (i) below 8 mM Im, the rate increased with concentration in a parabolic manner; (ii) above 20 mM, the rate leveled off, indicating a rate-limiting conformational step with lifetime approximately 1 s; and (iii) at Im concentrations above 100 mM, the rate substantially increased again, also parabolically. In contrast, binding of MeIm followed a simple hyperbolic concentration dependence. The temperature dependences of the binding and release kinetics of Im and MeIm were also measured and revealed very large activation parameters for all reactions. The complex concentration dependence of the Im binding rate is not consistent with the popular model for soluble c-type cytochromes in which exogenous ligand binding is preceded by spontaneous opening of the heme cleft, which becomes rate-limiting at high ligand concentrations. Instead, binding of ligand to the heme is explained by a model in which an initial and superficial binding facilitates access to the heme by disruption of hydrogen-bonded structures in the heme domain. For imidazole, two separate pathways of heme access are indicated by the distinct kinetics at low and high concentration. The structural basis for ligand entry to the heme cleft is discussed.
    Journal of Biological Chemistry 05/2010; 285(29):22522-31. · 4.65 Impact Factor
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    ABSTRACT: We have used imidazole (Im) and N-methylimidazole (MeIm) as probes of the heme-binding cavity of membrane-bound cytochrome (cyt) c(1) in detergent-solubilized bc(1) complex from Rhodobacter sphaeroides. Imidazole binding to cyt c(1) substantially lowers the midpoint potential of the heme and fully inhibits bc(1) complex activity. Temperature dependences showed that binding of Im (K(d) approximately 330 microM, 25 degrees C, pH 8) is enthalpically driven (DeltaH(0) = -56 kJ/mol, DeltaS(0) = -121 J/mol/K), whereas binding of MeIm is 30 times weaker (K(d) approximately 9.3 mM) and is entropically driven (DeltaH(0) = 47 kJ/mol, DeltaS(0)(o) = 197 J/mol/K). The large enthalpic and entropic contributions suggest significant structural and solvation changes in cyt c(1) triggered by ligand binding. Comparison of these results with those obtained previously for soluble cyts c and c(2) suggested that Im binding to cyt c(1) is assisted by formation of hydrogen bonds within the heme cleft. This was strongly supported by molecular dynamics simulations of Im adducts of cyts c, c(2), and c(1), which showed hydrogen bonds formed between the N(delta)H of Im and the cyt c(1) protein, or with a water molecule sequestered with the ligand in the heme cleft.
    Journal of Biological Chemistry 05/2010; 285(29):22513-21. · 4.65 Impact Factor
  • Erik W. Martin, Sergei Dikanov, Colin Wraight
    Biophysical Journal 01/2010; 98(3). · 3.67 Impact Factor
  • Erik W. Martin, Sergei Dikanov, Colin A. Wraight
    Biophysical Journal 01/2010; 98. · 3.67 Impact Factor

Publication Stats

1k Citations
320.17 Total Impact Points

Institutions

  • 1977–2014
    • University of Illinois, Urbana-Champaign
      • • Center for Biophysics and Computational Biology
      • • Department of Biochemistry
      • • Department of Plant Biology
      Urbana, Illinois, United States
  • 2008
    • University of Szeged
      • Institute of Physics
      Algyő, Csongrád, Hungary
  • 1999
    • Carl Zeiss AG
      Oberkochen, Baden-Württemberg, Germany
  • 1996
    • Academia Sinica
      T’ai-pei, Taipei, Taiwan
  • 1993
    • Moscow State Textile University
      Moskva, Moscow, Russia
  • 1987
    • Stanford University
      Palo Alto, California, United States