Retinal Binding to Apo-Gloeobacter Rhodopsin: The Role of pH and Retinal–Carotenoid Interaction

• Published in: The journal of physical chemistry. B Vol. 121; no. 48; pp. 10759 - 10769
• Main Authors: Jana, Sankar, Eliash, Tamar, Jung, Kwang-Hwan, Sheves, Mordechai
• Format: Journal Article
• Language: English
• Published: United States American Chemical Society 07-12-2017
• ISSN: 1520-6106; 1520-5207
• DOI: 10.1021/acs.jpcb.7b07523

Over the past few decades, the structure, functions, properties, and molecular mechanisms of retinal proteins have been studied extensively. The newly studied retinal protein Gloeobacter rhodopsin (gR) acts as a light-driven proton pump, transferring a proton from the cytoplasmic region to the extracellular region of a cell following light absorption. It was previously shown that gR can bind the carotenoid salinixanthin (sal). In the present study, we report the effect of pH on the binding of retinal to the apo-protein of gR, in the presence and absence of sal, to form the gR pigment. We found that binding at different pH levels reflects the titration of two different protein residues, one at the lower pK a 3.5 and another at the higher pK a 8.4, that affect the pigment’s formation. The maximum amount of pigment was formed at pH 5, both with and without the presence of sal. The introduction of sal accelerates the rate of pigment formation by a factor of 190. Furthermore, it is suggested that occupation of the binding site by the retinal chromophore induces protein conformational alterations which in turn affect the carotenoid conformation, which precedes the formation of the retinal–protein covalent bond. Our examination of synthetic retinal analogues in which the ring structure was modified revealed that, in the absence of sal, the retinal ring structure affects the rate of pigment formation and that the intact structure is needed for efficient pigment formation. However, the presence of sal abolishes this effect, and all-trans retinal and its modified ring analogues bind at a similar rate.