Method for lyophilizing brain proteolipid preparations that increases subsequent solubilization by detergents.
A frozen mixture of solubilized brain proteolipid proteins in chloroform-methanol is not sublimable in a vacuum. However, when 7 to 10 volumes of benzene were added to a chloroform-methanol solution containing 5 mg of proteolipid protein per ml, the proteolipid proteins remained in solution for a while and the frozen mixture was easily sublimated at 2mm Hg. Before the addition of benzene, higher concentrations of protein required the acidification of the medium to avoid precipitation of proteolipid proteins. In contrast to what happens when proteolipid proteins are obtained by the evaporation of the organic mixture at room temperature, the protein obtained by lyophilization was soluble in aqueous solutions of ionic and nonionic detergents. Sodium dodecyl sulfate at 0.6 to 0.7% concentration completely solubilized the proteolipid protein obtained by lyophilization. With the nonionic detergents Lubrol WX and Triton X-100, a solubilization between 50 and 65% was achieved. Sodium deoxycholate was practically ineffective. Triton X-100 showed selectivity in solubilizing certain proteins. The role of lipids in the solubilization of proteolipid proteins with detergents is discussed.
D000818:Animals; D001923:Brain Chemistry; D002417:Cattle; D003840:Deoxycholic Acid; D003902:Detergents; D005612:Freeze Drying / Q000379:methods; D017830:Octoxynol; D011092:Polyethylene Glycols; D011510:Proteolipids / Q000302:isolation & purification*; D012967:Sodium Dodecyl Sulfate; D012995:Solubility; D013501:Surface-Active Agents
D003902:Detergents; D011510:Proteolipids; D013501:Surface-Active Agents; D003840:Deoxycholic Acid; C017474:Lubrol WX; D012967:Sodium Dodecyl Sulfate; D011092:Polyethylene Glycols; D017830:Octoxynol