Separation of membrane proteins solubilized with a nondenaturing detergent and a high salt concentration by hydroxyapatite high-performance liquid chromatography. 1995

T Ichimura, and N Ikuta, and Y Uda, and T Horigome, and S Omata
Department of Biochemistry, Faculty of Science, Niigata University, Japan.

Ceramic hydroxyapatite high-performance liquid chromatography with a solvent containing high concentrations of salts was used for the separation of membrane proteins solubilized under nondenaturing and high salt conditions. The chromatographic conditions were optimized using sodium cholate as the detergent. By this method, most membrane proteins, prepared from rat liver rough microsomes, were effectively resolved from each other with a protein recovery of more than 90%. The method also allowed the single-step purification of the ribosome-binding protein, p34, from a microsomal membrane protein fraction. The good resolution with this method should be applicable to the isolation and characterization of a variety of membrane proteins on the analytical and semipreparative scales. With only the substitution of sodium cholate with other nondenaturing detergents, this method may also be applicable to the purification of membrane proteins requiring such nondenaturing detergents with retention of their biological activities.

UI MeSH Term Description Entries
D008565 Membrane Proteins Proteins which are found in membranes including cellular and intracellular membranes. They consist of two types, peripheral and integral proteins. They include most membrane-associated enzymes, antigenic proteins, transport proteins, and drug, hormone, and lectin receptors. Cell Membrane Protein,Cell Membrane Proteins,Cell Surface Protein,Cell Surface Proteins,Integral Membrane Proteins,Membrane-Associated Protein,Surface Protein,Surface Proteins,Integral Membrane Protein,Membrane Protein,Membrane-Associated Proteins,Membrane Associated Protein,Membrane Associated Proteins,Membrane Protein, Cell,Membrane Protein, Integral,Membrane Proteins, Integral,Protein, Cell Membrane,Protein, Cell Surface,Protein, Integral Membrane,Protein, Membrane,Protein, Membrane-Associated,Protein, Surface,Proteins, Cell Membrane,Proteins, Cell Surface,Proteins, Integral Membrane,Proteins, Membrane,Proteins, Membrane-Associated,Proteins, Surface,Surface Protein, Cell
D002793 Cholic Acids The 3 alpha,7 alpha,12 alpha-trihydroxy-5 beta-cholanic acid family of bile acids in man, usually conjugated with glycine or taurine. They act as detergents to solubilize fats for intestinal absorption, are reabsorbed by the small intestine, and are used as cholagogues and choleretics. Cholalic Acids,Acids, Cholalic,Acids, Cholic
D002851 Chromatography, High Pressure Liquid Liquid chromatographic techniques which feature high inlet pressures, high sensitivity, and high speed. Chromatography, High Performance Liquid,Chromatography, High Speed Liquid,Chromatography, Liquid, High Pressure,HPLC,High Performance Liquid Chromatography,High-Performance Liquid Chromatography,UPLC,Ultra Performance Liquid Chromatography,Chromatography, High-Performance Liquid,High-Performance Liquid Chromatographies,Liquid Chromatography, High-Performance
D003902 Detergents Purifying or cleansing agents, usually salts of long-chain aliphatic bases or acids, that exert cleansing (oil-dissolving) and antimicrobial effects through a surface action that depends on possessing both hydrophilic and hydrophobic properties. Cleansing Agents,Detergent Pods,Laundry Detergent Pods,Laundry Pods,Syndet,Synthetic Detergent,Agent, Cleansing,Agents, Cleansing,Cleansing Agent,Detergent,Detergent Pod,Detergent Pod, Laundry,Detergent Pods, Laundry,Detergent, Synthetic,Detergents, Synthetic,Laundry Detergent Pod,Laundry Pod,Pod, Detergent,Pod, Laundry,Pod, Laundry Detergent,Pods, Detergent,Pods, Laundry,Pods, Laundry Detergent,Synthetic Detergents
D017886 Durapatite The mineral component of bones and teeth; it has been used therapeutically as a prosthetic aid and in the prevention and treatment of osteoporosis. Calcium Hydroxyapatite,Hydroxyapatite,Hydroxylapatite,Alveograf,Calcitite,Interpore-200,Interpore-500,Osprovit,Ossein-Hydroxyapatite Compound,Ossopan,Osteogen,Periograf,Hydroxyapatite, Calcium,Interpore 200,Interpore 500,Interpore200,Interpore500,Ossein Hydroxyapatite Compound
D019826 Cholic Acid A major primary bile acid produced in the liver and usually conjugated with glycine or taurine. It facilitates fat absorption and cholesterol excretion. Cholbam

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