Binding of alkyl polyglucoside surfactants to bacteriorhodopsin and its relation to protein stability.
The binding of alkyl polyglucoside surfactants to the integral membrane protein bacteriorhodopsin (BR) and the formation of protein-surfactant complexes are investigated by sedimentation equilibrium via analytical ultracentrifugation and by small-angle neutron scattering (SANS). Contrast variation techniques in SANS enable measurement of the composition of the protein-surfactant complexes and determination of the thickness of the surfactant shell bound to the protein. The results indicate that alkyl polyglucosides can bind to BR as single surfactant layers or as a thicker shell. The thickness of the surfactant shell increases with increasing surfactant tail length, and it is generally unrelated to the aggregation number of the micelles even for a small and predominantly hydrophobic membrane protein such as BR. The aggregation numbers determined by sedimentation equilibrium methods match those measured by SANS, which also allows reconstruction of the shape of the protein-detergent complex. When the surfactant is present as a single layer, the BR loses activity, as measured by absorption spectroscopy, more quickly than it does when the surfactant forms a thicker shell.
Santonicola
M Gabriella
MG
Center for Molecular and Engineering Thermodynamics, Department of Chemical Engineering, University of Delaware, Newark, Delaware 19716-3119, USA.
Lenhoff
Abraham M
AM
Kaler
Eric W
EW
D000042:Absorption / Q000187:drug effects; D001436:Bacteriorhodopsins / Q000378:metabolism*; D005960:Glucosides / Q000378:metabolism*; D019617:Halobacterium salinarum; D008823:Micelles; D008956:Models, Chemical; D011134:Polysaccharides / Q000378:metabolism* / Q000494:pharmacology*; D011485:Protein Binding; D012995:Solubility; D013056:Spectrophotometry, Ultraviolet; D013501:Surface-Active Agents / Q000378:metabolism* / Q000494:pharmacology*
D005960:Glucosides; D008823:Micelles; D011134:Polysaccharides; D013501:Surface-Active Agents; D001436:Bacteriorhodopsins