The interaction of Triton X-100 with purple membranes. Detergent binding, spectral changes and membrane solubilization.
The interaction of the non-ionic surfactant Triton X-100 with Halobacterium purple membranes has been examined at sublytic and lytic surfactant concentrations. These membranes present a number of important peculiarities in their behaviour towards the surfactant. Although solubilization is a very slow process, with a half-time of the order of hours, detergent binding appears to occur at the same fast rate as that found in other membranes. Lipids are solubilized more easily than proteins, so that hardly any protein is solubilized at surfactant concentrations at which about 75% of the lipid is in the form of detergent-mixed micelles; once started, protein solubilization takes place within a narrow range of surfactant concentrations. Retinal provides a built-in probe to monitor detergent-induced conformational changes by spectroscopy in the visible range. No spectral variation is detected at the prelytic stage, i.e. when detergent is incorporated into the membrane in monomeric form. Membrane disruption is accompanied by a blue shift in the absorption maximum, retinal isomerization (from all-trans to 13-cis), and a decrease in specific absorbance (bleaching). Increasing detergent concentrations after solubilization is completed do not produce further shifts in the spectral maximum, but the specific absorbance is progressively decreased. It is shown that Triton X-100 has a complex effect on the retinal chromophore, modifying its configuration and microenvironment (changes in maximum wavelength) and promoting hydrolysis of the retinal-bacteriorhopsin Schiff's base (bleaching).
González-Mañas
J M
JM
Department of Biochemistry, University of the Basque Country, Bilbao, Spain.
Virto
M D
MD
Gurtubay
J I
JI
Goñi
F M
FM
D000220:Adaptation, Biological; D001436:Bacteriorhodopsins / Q000032:analysis*; D001665:Binding Sites; D003902:Detergents; D006217:Halobacterium / Q000032:analysis; D007536:Isomerism; D008027:Light; D017830:Octoxynol; D011092:Polyethylene Glycols / Q000032:analysis*; D011487:Protein Conformation; D012172:Retinaldehyde / Q000032:analysis; D012995:Solubility
D003902:Detergents; D011092:Polyethylene Glycols; D001436:Bacteriorhodopsins; D017830:Octoxynol; D012172:Retinaldehyde