Triplet-minus-singlet absorbance difference spectra of reaction centers of Rhodopseudomonas sphaeroides R-26 in the temperature range 24-290 K measured by Magneto-Optical Difference Spectroscopy (MODS).
The recently developed technique of Magneto-Optical Difference Spectroscopy (MODS) [10] has been applied to reaction centers (RC) of the photosynthetic bacterium Rhodopseudomonas sphaeroides R-26. Absorbance changes induced by a magnetic field are measured as a function of wavelength yielding the triplet-minus-singlet (T-S) absorbance difference spectrum. (T-S) spectra thus obtained have been measured from 24-290 K. Going from low to high temperature the (T-S) spectra show the following features: (a) A rapid decrease of positive absorption bands at 809 and 819 nm. (b) A slow appearance of a band shift at 798 nm. (c) A shift of the peak wavelength of the Qy absorbance band of the primary donor P-860 from 992 to 861 nm, and of its Qx band from 603 to 600 nm. The spectra at 24, 66, 116, and 290 K have been analyzed by Gaussian deconvolution. The 800 nm region of the spectrum at 24 K can be decomposed in a combination of two band shifts and an appearing band. The temperature dependence of the spectra in this region is well explained by spectral broadening of the two shifting bands combined with a decrease in intensity of the appearing band when the temperature increases.The two shifting bands in the 800 nm region are identified as the two bands at 803 and 813 nm which together make up the 800 nm band in the absorption spectrum and are assigned to the two accessory RC bacteriochlorophylls (BChls). The band shift of the 813 nm pigment is appreciably larger than that of the 803 nm pigment. The appearing band at 808 nm is attributed to monomeric absorption of (3)P-860, the triplet state being localized on one BChl.We find no evidence for admixture of a charge transfer (CT) state of (3)P-860 with one of the accessory BChls at higher temperature.
Department of Biophysies, Huygens Laboratory of the State University, P.O. Box 9504, 2300 RA, Leiden, The Netherlands.
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