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Method for Lyophilizing Brain Proteolipid Preparation That Increases Subsequent Solubilization by Detergent
Author(s) -
Aguilar J. S.,
Cózar M. de,
Criado M.,
Monreal J.
Publication year - 1982
Publication title -
journal of neurochemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.75
H-Index - 229
eISSN - 1471-4159
pISSN - 0022-3042
DOI - 10.1111/j.1471-4159.1982.tb08010.x
Subject(s) - chemistry , chloroform , solubilization , chromatography , benzene , sodium dodecyl sulfate , proteolipid protein 1 , methanol , biochemistry , organic chemistry , myelin basic protein , myelin , neuroscience , biology , central nervous system
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 2 mm 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.