The hydrogen-bonding network of water molecules and the peptide backbone in the region connecting Asp83, Gly120, and Glu113 in bovine rhodopsin.

Difference Fourier transform infrared spectra were recorded between mutants of rhodopsin and their batho products. The pigments studied were single and combined mutants of intramembrane residues of bovine rhodopsin: Asp83, Glu113, Gly120, Gly121, and Glu122. Previous studies [Nagata, T., Terakita, A., Kandori, H., Kojima, D., Shichida, Y., and Maeda, A. (1997) Biochemistry 36, 6164-6170] showed that one of the water molecules which undergoes structural changes in this process forms hydrogen bonds with Glu113 and the Schiff base, and that another water molecule is linked to this structure through the peptide backbone. The present results show that this water molecule is located at the place that is affected by the replacements of Asp83 and Gly120 but only slightly by Gly121 and not at all by Glu122. Asp83 and Gly120 are close to each other, in view of the observations that the carboxylic C=O stretching vibration of Asp83 is affected by the G120A replacement and that each replacement affects the common peptide carbonyl groups. Our results suggest that these residues in the middle of helices B and C are linked-through a hydrogen-bonding network composed of water and the peptide backbone-with the region around Glu113.