Subfractions of membranes from calf brain synaptosomes obtained and studied by liquid-liquid partitioning.
Synaptosomes isolated from calf brain cortex were lysed and fragmented by Yeda press treatment. The obtained membranes have previously been fractionated in a counter-current distribution process using a liquid-liquid two-phase system consisting of water, dextran, Ficoll and poly(ethylene glycol) [J. Chromatogr., 358 (1986) 147]. Using the fact that there are discrete membrane populations, a rapid preparative method for isolation of the two main fractions is presented in the present work, as well as a subfractionation of one of them using liquid-liquid extraction with dextran-bound Procion yellow HE-3G. The content of several membrane constituents, i.e. protein, acetylcholinesterase, succinate dehydrogenase and ATPase, as well as opiate binding, were determined for the three fractions. Counter-current distribution of the fractions elucidates their heterogeneity and the effectiveness of the purification.
Muiño Blanco
M T
MT
Department of Biochemistry, University of Lund, Sweden.
Alvaro Cebrian
J
J
Olde
B
B
Johansson
G
G
D000110:Acetylcholinesterase / Q000032:analysis; D000251:Adenosine Triphosphatases / Q000032:analysis; D000818:Animals; D001923:Brain Chemistry; D002417:Cattle; D002458:Cell Fractionation; D002540:Cerebral Cortex / Q000737:chemistry* / Q000166:cytology / Q000648:ultrastructure; D002853:Chromatography, Liquid / Q000379:methods*; D003911:Dextrans; D005026:Ethylene Glycols; D005362:Ficoll; D008565:Membrane Proteins / Q000032:analysis; D013385:Succinate Dehydrogenase / Q000032:analysis; D013570:Synaptic Membranes / Q000737:chemistry*; D013574:Synaptosomes / Q000737:chemistry* / Q000648:ultrastructure; D014227:Triazines
D003911:Dextrans; D005026:Ethylene Glycols; D008565:Membrane Proteins; D014227:Triazines; D005362:Ficoll; C009794:procion yellow; D013385:Succinate Dehydrogenase; D000110:Acetylcholinesterase; D000251:Adenosine Triphosphatases