Deoxylysolecithin and a new biphenyl detergent as solubilizing agents for bovine rhodopsin. Functional test by formation of metarhodopsin II and binding of G-protein.
The protein-detergent interaction in rhodopsin-detergent micelles has been investigated by using formation of metarhodopsin II (MII) as a monitor. Two detergents of different structural rigidity have been applied. One of them is [3-(lauroyloxy)propyl]phosphorylcholine, which has a high conformational flexibility in its hydrophobic moiety like most of the known detergents for rhodopsin. This deoxylysolecithin was originally designed as a detergent for membrane proteins by Weltzien [Weltzien, H. U. (1979) Biochim. Biophys. Acta 559, 259-287]. The other detergent, which is highly rigid in its hydrophobic part, has been developed for this study. It consists of a biphenyl derivative and a hydrophilic octaethylene oxide group. Both the formation kinetics of MII and the position of its equilibrium with its tautomeric form, metarhodopsin I (MI), strongly differed in the deoxylysolecithin and biphenyl detergent. Deoxylysolecithin caused very fast MII formation and shifted the equilibrium strongly to MII, like other detergents with alkyl chains as the hydrophobic part. In the biphenyl detergent, however, formation of MII was slow and the MI/MII equilibrium similar to that in the native system. For rhodopsin reconstituted in lipid bilayers, normal MII formation requires a well-adjusted fluidity of the hydrocarbon environment of the protein [Baldwin, P. A., & Hubbell, W. L. (1984) Biochemistry 24, 2633-2639], which was explained by an appropriate interfacial pressure at the protein-lipid interface. Extension of this concept would indicate that in the micellar core a degree of fluidity comparable to that of the disk membrane is just achieved with the highly rigid biphenyl structure.(ABSTRACT TRUNCATED AT 250 WORDS)
D000818:Animals; D001713:Biphenyl Compounds / Q000138:chemical synthesis; D002417:Cattle; D003902:Detergents / Q000138:chemical synthesis; D019204:GTP-Binding Proteins / Q000378:metabolism*; D007202:Indicators and Reagents; D008244:Lysophosphatidylcholines; D009682:Magnetic Resonance Spectroscopy; D011485:Protein Binding; D012168:Retinal Pigments / Q000378:metabolism*; D012243:Rhodopsin / Q000031:analogs & derivatives / Q000302:isolation & purification / Q000378:metabolism*; D012374:Rod Cell Outer Segment / Q000378:metabolism; D012995:Solubility; D013501:Surface-Active Agents / Q000138:chemical synthesis
D001713:Biphenyl Compounds; D003902:Detergents; D007202:Indicators and Reagents; D008244:Lysophosphatidylcholines; D012168:Retinal Pigments; D013501:Surface-Active Agents; C013110:metarhodopsins; D012243:Rhodopsin; D019204:GTP-Binding Proteins