The mechanism of detergent solubilization of liposomes and protein-containing membranes

被引:266
作者
Kragh-Hansen, U
le Maire, M
Moller, JV
机构
[1] Aarhus Univ, Dept Biophys, DK-8000 Aarhus C, Denmark
[2] Aarhus Univ, Dept Med Biochem, DK-8000 Aarhus C, Denmark
[3] CEA, Dept Biol Cellulaire & Mol, Sect Biophys Prot & Membranes, F-91191 Gif Sur Yvette, France
[4] Ctr Etud Saclay, CNRS, URA 2096, F-91191 Gif Sur Yvette, France
关键词
D O I
10.1016/S0006-3495(98)77735-5
中图分类号
Q6 [生物物理学];
学科分类号
071011 ;
摘要
The present study explores intermediate stages in detergent solubilization of liposomes and Ca2+-ATPase membranes by sodium dodecyl sulfate (SDS) and medium-sized (similar to C-12) 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.
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页码:2932 / 2946
页数:15
相关论文
共 64 条
[1]   PERTURBATION OF THE STRUCTURE AND FUNCTION OF A MEMBRANOUS CA-2+-ATPASE BY NON-SOLUBILIZING CONCENTRATIONS OF A NON-IONIC DETERGENT [J].
ANDERSEN, JP ;
LEMAIRE, M ;
KRAGHHANSEN, U ;
CHAMPEIL, P ;
MOLLER, JV .
EUROPEAN JOURNAL OF BIOCHEMISTRY, 1983, 134 (02) :205-214
[2]  
BARTLETT GR, 1959, J BIOL CHEM, V234, P466
[3]   PHOSPHOLIPID FATTY ACYL CHAIN ASYMMETRY IN THE MEMBRANE BILAYER OF ISOLATED SKELETAL-MUSCLE SARCOPLASMIC-RETICULUM [J].
BICK, RJ ;
VANWINKLE, WB ;
TATE, CA ;
ENTMAN, ML ;
BLASIE, JK ;
HERBETTE, LG .
BIOCHEMISTRY, 1987, 26 (15) :4831-4836
[4]   Dimer to monomer conversion of the cytochrome b(6)f complex - Causes and consequences [J].
Breyton, C ;
Tribet, C ;
Olive, J ;
Dubacq, JP ;
Popot, JL .
JOURNAL OF BIOLOGICAL CHEMISTRY, 1997, 272 (35) :21892-21900
[5]  
Cornelius F., 1995, HDB NONMEDICAL APPL, V2, P219
[6]   CRYSTAL-STRUCTURES EXPLAIN FUNCTIONAL-PROPERTIES OF 2 ESCHERICHIA-COLI PORINS [J].
COWAN, SW ;
SCHIRMER, T ;
RUMMEL, G ;
STEIERT, M ;
GHOSH, R ;
PAUPTIT, RA ;
JANSONIUS, JN ;
ROSENBUSCH, JP .
NATURE, 1992, 358 (6389) :727-733
[7]   2-D STRUCTURE OF THE NEUROSPORA-CRASSA PLASMA-MEMBRANE ATPASE AS DETERMINED BY ELECTRON CRYOMICROSCOPY [J].
CYRKLAFF, M ;
AUER, M ;
KUHLBRANDT, W ;
SCARBOROUGH, GA .
EMBO JOURNAL, 1995, 14 (09) :1854-1857
[8]   MEMBRANE SOLUBILIZATION BY DETERGENT - USE OF BROMINATED PHOSPHOLIPIDS TO EVALUATE THE DETERGENT-INDUCED CHANGES IN CA-2+-ATPASE LIPID INTERACTION [J].
DEFORESTA, B ;
LEMAIRE, M ;
ORLOWSKI, S ;
CHAMPEIL, P ;
LUND, S ;
MOLLER, JV ;
MICHELANGELI, F ;
LEE, AG .
BIOCHEMISTRY, 1989, 28 (06) :2558-2567
[9]   KINETIC CHARACTERIZATION OF THE PERTURBATION BY DODECYLMALTOSIDE OF SARCOPLASMIC-RETICULUM CA2+-ATPASE [J].
DEFORESTA, B ;
HENAO, F ;
CHAMPEIL, P .
EUROPEAN JOURNAL OF BIOCHEMISTRY, 1992, 209 (03) :1023-1034
[10]  
DEFORESTA B, 1994, EUR J BIOCHEM, V223, P359