New zwitterionic detergents improve the analysis of membrane proteins by two-dimensional electrophoresis.
Severe quantitative loss of protein is often observed in high-resolution two-dimensional electrophoresis of membrane proteins, while the resolution is usually not affected. To improve the solubility of proteins in this technique, we tested denaturing cocktails containing various detergents and chaotropes. Best results were obtained with a denaturing solution containing urea, thiourea, and zwitterionic detergents, synthesized for this purpose. Among the dozen detergents synthesized and tested, amidosulfobetaines with an alkyl tail containing 14-16 carbons proved most efficient, solubilizing previously undetected membrane proteins.
Chevallet
M
M
CEA, Laboratoire de Bioénergétique Cellulaire et Pathologique, Grenoble, France.
Santoni
V
V
Poinas
A
A
Rouquié
D
D
Fuchs
A
A
Kieffer
S
S
Rossignol
M
M
Lunardi
J
J
Garin
J
J
Rabilloud
T
T
D000818:Animals; D017360:Arabidopsis; D015153:Blotting, Western; D002417:Cattle; D003902:Detergents / Q000737:chemistry; D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods; D008565:Membrane Proteins / Q000032:analysis*; D015394:Molecular Structure; D009504:Neutrophils / Q000737:chemistry*; D010940:Plant Proteins / Q000032:analysis*
D003902:Detergents; D008565:Membrane Proteins; D010940:Plant Proteins