Detergent effects on enzyme activity and solubilization of lipid bilayer membranes.
Over 50 detergents were tested to establish which would be most effective in releasing proteins from membrane-bounded compartments without denaturing them. Various concentrations each of detergent were tested for two activities: (1) solubilization of egg phospholipid liposomes as measured by reduction of turbidity and (2) effect of detergent concentration on the activities of soluble, hydrolytic enzymes. Those detergents most effective in solubilizing 0.2% lipid and least detrimental to enzymes were five pure, synthetic compounds recently introduced: CHAPS, CHAPSO, Zwittergents 310 and 312, and octylglucoside. Industrial detergents were generally much inferior, insofar as they solubilized membranes inefficiently and/or inactivated certain hydrolytic enzymes readily. The five detergents were characterized by (a) an unusually high critical micelle concentration and (b) a preference for forming mixed micelles with lipids instead of forming pure micelles, as indicated by an ability to solubilize lipid at concentrations of detergent significantly below the critical micelle concentration. This characteristic permits solubilization of high concentrations of membrane below the critical micelle concentration of the detergent so that protein denaturation is minimized. A generally applicable guideline that emerged from this study is that detergents should be used at approximately their critical micelle concentration which should not be exceeded by the concentration of membrane. Similar considerations should apply to the use of detergents in purifying and reconstituting intrinsic membrane proteins.
D000469:Alkaline Phosphatase / Q000378:metabolism*; D003902:Detergents / Q000494:pharmacology*; D004950:Esterases / Q000378:metabolism*; D007700:Kinetics; D008051:Lipid Bilayers; D010713:Phosphatidylcholines; D010944:Plants / Q000201:enzymology; D012995:Solubility; D013429:Sulfatases / Q000378:metabolism*; D013501:Surface-Active Agents / Q000494:pharmacology*; D014908:Triticum / Q000201:enzymology
D003902:Detergents; D008051:Lipid Bilayers; D010713:Phosphatidylcholines; D013501:Surface-Active Agents; D004950:Esterases; D000469:Alkaline Phosphatase; D013429:Sulfatases