Abstract
Co-reconstitution of photosynthetic reaction centers (RCs) with bc1 complexes, both from Rb. Sphaeroides, has been investigated using surface-enhanced IR-absorption spectroscopy (SEIRAS) in the ATR configuration. Surface-enhancement is achieved by a thin nano-structured gold film present on the surface of an ATR crystal. The proteins were simultaneously immobilized as a monolayer on the gold film via His-tag technology and subsequently co-reconstituted into a lipid bilayer by in-situ dialysis. Steady-state light-minus-dark absorbance spectra were recorded, indicating the excitation of the special pair as well as alterations of quinone/quinol species. Band assignment was facilitated by co-reconstitution of liposoluble Q10 into the lipid phase. Changes of the SEIRA spectra were observed, particularly after addition of cytochrome c, indicating the interaction of the RC with the bc1 complex via the quinone/quinol pool.
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