The mechanism of detergent solubilization of liposomes and protein-containing membranes.
The present study explores intermediate stages in detergent solubilization of liposomes and Ca2+-ATPase membranes by sodium dodecyl sulfate (SDS) and medium-sized ( approximately C12) nonionic detergents. In all cases detergent partitioning in the membranes precedes cooperative binding and solubilization, which is facilitated by exposure to detergent micelles. Nonionic detergents predominantly interact with the lipid component of Ca2+-ATPase membranes below the CMC (critical micellar concentration), whereas SDS extracts Ca2+-ATPase before solubilization of lipid. At the transition to cooperative binding, n-dodecyl octaethylene glycol monoether (C12E8), Triton X-100, and dodecyldimethylamine oxide induce fusion of small unilamellar liposomes to larger vesicles before solubilization. Solubilization of Ca2+-ATPase membranes is accompanied by membrane fragmentation and aggregation rather than vesicle fusion. Detergents with strongly hydrophilic heads (SDS and beta-D-dodecylmaltoside) only very slowly solubilize liposomal membranes and do not cause liposome fusion. These properties are correlated with a slow bilayer flip-flop. Our data suggest that detergent solubilization proceeds by a combination of 1) a transbilayer attack, following flip-flop of detergent molecules across the lipid bilayer, and 2) extraction of membrane components directly by detergent micelles. The present study should help in the design of efficient solubilization protocols, accomplishing the often delicate balance between preserving functional properties of detergent sensitive membrane proteins and minimizing secondary aggregation and lipid content.
Kragh-Hansen
U
U
Department of Medical Biochemistry, University of Aarhus, DK-8000 Aarhus C, Denmark.
le Maire
M
M
Møller
J V
JV
D000818:Animals; D055592:Biophysical Phenomena; D001703:Biophysics; D000252:Calcium-Transporting ATPases / Q000302:isolation & purification; D003902:Detergents; D007700:Kinetics; D008051:Lipid Bilayers; D008081:Liposomes; D008565:Membrane Proteins / Q000302:isolation & purification*; D008823:Micelles; D010713:Phosphatidylcholines; D011485:Protein Binding; D011817:Rabbits; D012967:Sodium Dodecyl Sulfate; D012995:Solubility; D013816:Thermodynamics
D003902:Detergents; D008051:Lipid Bilayers; D008081:Liposomes; D008565:Membrane Proteins; D008823:Micelles; D010713:Phosphatidylcholines; D012967:Sodium Dodecyl Sulfate; D000252:Calcium-Transporting ATPases; C017251:1,2-oleoylphosphatidylcholine