On the mechanism of bacteriorhodopsin solubilization by surfactants.
Purple membrane bacteriorhodopsin can be easily solubilized by Triton X-100 and other detergents, but not by deoxycholate. In order to understand this behavior, we have examined the effects of a variety of surfactants. We show that detergents containing the cholane ring (cholate, taurocholate, 3[(3-cholamidopropyl)diethyl-ammonio]propanesulfonic acid...) are virtually unable to solubilize native bacteriorhodopsin. However, when the protein is reconstituted in dimyristoyl phosphatidylcholine and solubilization is assayed at a temperature such that bacteriorhodopsin is in the form of monomers, solubilization by cholane detergents does occur. We propose that steric factors prevent access of the rigid planar surfactant molecules to the hydrophobic protein regions. These are perhaps located in the monomer-monomer interface, whose solvation by surfactants is essential for solubilization to occur. We note that the capacity of some detergents to solubilize bacteriorhodopsin is always associated within the same range of surfactant concentrations with bleaching (partial or total) of the protein chromophore. The detergent-induced bleaching is at least partially reversible, suggesting that free retinal remains associated to some membrane components. While some surfactant molecules remain tightly bound to the membrane protein, cholane detergents can be completely removed from bacteriorhodopsin. Our results indicate that a structure-function relationship exists for detergents applied to the solubilization of bacteriorhodopsin.
del Río
E
E
Department of Biochemistry, University of the Basque Country, Bilbao, Spain.
González-Mañas
J M
JM
Gurtubay
J I
JI
Goñi
F M
FM
D001436:Bacteriorhodopsins / Q000378:metabolism*; D002757:Cholanes; D003902:Detergents; D008565:Membrane Proteins / Q000378:metabolism*; D010743:Phospholipids; D012995:Solubility; D013056:Spectrophotometry, Ultraviolet; D013501:Surface-Active Agents / Q000378:metabolism*
D002757:Cholanes; D003902:Detergents; D008565:Membrane Proteins; D010743:Phospholipids; D013501:Surface-Active Agents; D001436:Bacteriorhodopsins