Abstract
Bacteriorhodopsin (bR), converted by deionization to the blue form was reconstituted to the active purple membrane by the addition of Fe2+ or Fe3+ ions. 57Fe Mossbauer spectra of these samples were measured at different pH values (pH 3.9, pH 5.0 and pH 7.0) and at temperatures ranging from 4 K to 300 K. The hyperfine parameters reveal two iron environments with oxygen atoms in the neighbourhood of iron. Iron type 1 is in the 3+ high spin state. It is bound to acid side chains of the protein and/or the phosphate groups of the lipids. Iron type 2 is in the 2+ high spin state and is linked to carboxy groups of the protein in a rather unspecific way. Dynamics as measured by Mossbauer spectroscopy show that the purple membrane becomes flexible only above 220 K. At the interface between membrane and bulk water the mobility is comparable to that of proteins with hydrophilic surfaces. The photocycle of Fe 3+-bR is slowed down compared to native bR. 3–5 Fe3+/bR are sufficient to inhibit the photocycle turnover by one order of magnitude. This specific effect is also found with Cr3+, though it is less pronounced. Mössbauer spectra of Fe3+-bR at 4 K reveal that iron nuclei are spin-coupled, indicating their close spatial proximity. It is proposed that iron trinuclear clusters interact with the proton uptake site of bR.
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Engelhard, M., Kohl, K.D., Müller, K.H. et al. The photocycle and the structure of iron containing bacteriorhodopsin —a kinetic and Mössbauer spectroscopy investigation. Eur Biophys J 19, 11–18 (1990). https://doi.org/10.1007/BF00223568
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DOI: https://doi.org/10.1007/BF00223568