Evidence that Ala M260 is hydrogen-bonded to the reduced primary acceptor quinone QA−. in reaction centers of Rb. sphaeroides

The binding of the primary quinone anion radical Q(A)(-.) in reaction centers (RCs) of Rhodobacter (Rb.) sphaeroides species was investigated with electron spin echo envelope modulation (ESEEM). The ESEEM spectra, at two microwave frequencies, of Zn-substituted RCs of Rb. sphaeroides R26 showed interactions of the unpaired electron of Q(A)(-.) with two nitrogen nuclei in the protein matrix. From an analysis of the experimental data the nitrogen nuclear quadrupole resonance parameters were determined: e(2)qQ/h = 1.52 MHz, eta = 0.82 and e(2)qQ/h = 3.04 MHz, eta = 0.66, which are assigned to the N-14(8(1))-H group of His M219 and the peptide N-14 of Ala M260, The ESEEM spectrum of Q(A)(-.) in reaction centers of the Rb, sphaeroides mutant W(M252)Y shows that the nitrogen of Trp M252 is not interacting with Q(A)(-.), that of the mutant H(M266)C shows that manipulating the Fe binding site strongly affects the Q(A)-binding site

QA binding in reaction centers of the photosynthetic purple bacterium rhodobacter sphaeroides R26 investigated with electron spin polarization spectroscopy

The relation between quinone (QA) binding and electron transport in reaction centers (RCs) of photosynthetic purple bacteria is investigated, using electron spin polarization (ESP) X-band (9 GHz) EPR as a tool to probe for structural changes resulting from charge separation and stabilization and from replacing the native QA molecule with other quinones. We present a study of possible changes in Q(A)-binding that might be responsible for the remarkably prolonged Lifetime of the charge-separated state at cryogenic temperatures for RCs of Rhodobacter sphaeroides R26 cooled under illumination [Kleinfeld, D., et al. (1984) Biochemistry 23, 5780-5786]. It is shown that this effect is not caused by a major reorientation of the chromophores. Furthermore, we studied the effects of structurally different quinones functioning as primary electron acceptor in different purple bacteria. With simulations of ESP X-band spectra of the spin-polarized secondary radical pair P.(+)Q(A)(.-) in menaquinone-reconstituted, Zn2+-substituted RCs of Rb. sphaeroides R26, we show that quinone reconstitution is highly selective for site and orientation. Furthermore, we find that a very small exchange interaction between P-.+ and Q(A)(.-) (\J(PQ)\ similar to 1 mu T) is needed to account accurately for the observed relative line intensities at X-band, without affecting the accuracy of the simulations of reported ESP K-band spectra [Fuchsle, G., et al. (1993) Biochim. Biophys. Acta 1142, 23-35; Van der Est, A., et al. (1993) Chem. Phys. Lett. 212, 561-568]. This pronounced influence of small values for J(PQ) On the X-band ESP line shape results from cancellation effects of absorptive and emissive contributions to the spectrum, such that small shifts can be observed. The exchange interaction has opposite sign for the native, ubiquinone-containing RC [viz. J(P.UQ) (-0 8 +/- 0.2) mu T] and the menaquinone-substituted RC [J(P.MK) (+0.3 +/- 0.2)mu T]. The implications of these observations for electron-transport theory are discussed

Control of radical pair lifetimes by microwave irradiation: application to photosynthetic reaction centres

Radicals produced by illumination or ionizing radiation are often produced in pairs, which quickly decay by recombination or by diffusion and subsequent reactions. For maximizing the yield of products, and for facilitating the study of reaction pathways, it is desirable to minimize the probability of radical pair recombination. We present a way of controlling the radical pair lifetime through the application of a pulse of resonant microwaves in the presence of a magnetic field. Herewith, two radical pair triplet states are coherently populated, from which the pair cannot recombine directly to the singlet ground state because of spin conservation. We illustrate the method with a photosynthetic photochemical reaction, where we have achieved an increase in the radical pair lifetime of up to two orders of magnitude

Effect of bicarbonate on the S2 multiline EPR signal of the oxygen-evolving complex in photosystem II membrane fragments

Removal of bicarbonate from spinach photosystem II BBY particles by means of washing in a CO2-free medium results in the loss of their capability to accumulate the S2 multiline EPR signal upon continuous illumination at 190 K. Addition of 1 mM NaHCO3 before illumination leads to a 50-60% restoration of the multiline signal. Similarly, in BBY particles depleted of Mn by treatment with 1 mM Tris-HCl (pH 8.0) and 0.5 M MgCl2, re-addition of MnCl2 in the presence of 1 mM NaHCO3 results in a partial restoration (~ 30%) of the S2 multiline EPR signal of the Mn cluster, while in the absence of NaHCO3 no restoration is observed. The results provide further evidence that bicarbonate is essential for maintaining the Mn-containing oxygen-evolving complex of PS II in a functionally active form.Peer Reviewe