Crystallographic characterization by X-ray diffraction of the M-intermediate from the photo-cycle of bacteriorhodopsin at room temperature

The structure of the M-intermediate appearing in the photo-cycle of bacteriorhodopsin was studied with X-ray diffraction techniques at room temperature. The lifetime of the M-intermediate was prolonged by treatment with an arginine solution at alkaline pH (Nakasako et al.,FEBS Lett. 254, 211-214). The diffraction profile of membranes which had accumulated the M-intermediate had small but significant differences in the intensities of Bragg reflections and the lattice constant in comparison with that of membranes having trans-bacteriorhodopsin. Diffraction intensities were carefully evaluated and the structural changes during the formation of the intermediate were evaluated with difference Fourier analysis. We could find structural changes around helices G and B.