title: Influence of iron-removal procedures on sequential electron transfer in photosynthetic bacterial reaction centers studied by transient EPR spectroscopy.
abstract: Electron spin polarized electron paramagentic resonance (ESP EPR) spectra were obtained with deuterated iron-removed photosynthetic bacterial reaction centers (RCs) to specifically investigate the effect of the rate of primary charge separation, metal-site occupancy, and H-subunit content on the observed P865+QA- charge-separated state. Fe-removed and Zn-substituted RCs from Rb. sphaeroides R-26 were prepared by refined procedures, and specific electron transfer rates (kQ) from the intermediate acceptor H- to the primary acceptor QA of (200 ps)-1 vs (3-6 ns)-1 were observed. Correlation of the transient EPR and optical results shows that the observed slow kQ rate in Fe-removed RCs is H-subunit-independent, and, in some cases, independent of Fe-site occupancy as Zn2+ substitution does not ensure retention of the native kQ. In addition, shifts in the optical spectrum of P865 and differences in the high-field region of the Q-band ESP spectrum for Fe-removed RCs with slow kQ indicate possible structural changes near P865. The experimental X-band and Q-band spin-polarized EPR spectra for deuterated Fe-removed RCs where kQ is at least 15-fold slower at room temperature than the (200 ps)-1 rate observed for native Fe-containing RCs have different relative amplitudes and small g-value shifts compared to the spectra of Zn-RCs which have a kQ unchanged from native RCs. These differences reflect the trends in polarization predicted from the sequential electron transfer polarization (SETP) model [Morris et al. (1995) J. Phys. Chem. 99, 3854-3866; Tang et al. (1996) Chem. Phys. Lett. 253, 293-298]. Thus, SETP modeling of these highly resolved ESP spectra obtained with well-characterized proteins will provide definitive information about any light-induced structural changes of P865, H, and QA that occur upon formation of the P865+QA- charge-separated state.
authors.affiliation: Chemistry Division, Argonne National Laboratory, Argonne, Illinois 60439, USA.
authors.forename: L M
authors.initials: LM
authors.lastname: Utschig
authors.forename: S R
authors.initials: SR
authors.lastname: Greenfield
authors.forename: J
authors.initials: J
authors.lastname: Tang
authors.forename: P D
authors.initials: PD
authors.lastname: Laible
authors.forename: M C
authors.initials: MC
authors.lastname: Thurnauer
mesh_terms: D004578:Electron Spin Resonance Spectroscopy; D004579:Electron Transport; D007501:Iron / Q000032:analysis / Q000378:metabolism*; D017879:Isothiocyanates / Q000378:metabolism / Q000494:pharmacology; D007700:Kinetics; D008027:Light; D008345:Manganese / Q000032:analysis; D045322:Photosynthetic Reaction Center Complex Proteins / Q000378:metabolism*; D012242:Rhodobacter sphaeroides / Q000378:metabolism*; D013053:Spectrophotometry; D015032:Zinc / Q000032:analysis
chemical_list: D017879:Isothiocyanates; D045322:Photosynthetic Reaction Center Complex Proteins; D008345:Manganese; D007501:Iron; D015032:Zinc