Effect of dextran- and poly(ethylene glycol)-bound procion yellow HE-3G on the partition of membranes from calf brain synaptosomes within an aqueous two-phase system.
Membranes obtained by lysis and Yeda-press treatment of synaptosomes (nerve endings) from calf brain cortex have been partitioned within the aqueous phases (and the interface between them) of a Ficoll-dextran-poly(ethylene glycol)-water two-phase system. By introducing the dye Procion yellow HE-3G in the upper phase, bound to poly(ethylene glycol), or in the lower phase, bound to dextran, the partition of the membranes could be strongly affected. The influence on the partition was more pronounced when the dye was bound to dextran. By using a number of two-phase systems in a counter-current distribution process, it was shown that the membrane preparation was inhomogeneous and that the fractions obtained differed in their contents of acetylcholinesterase, succinate dehydrogenase and ATPase. The affinity partitioning effect depended strongly on the concentration of polymer-bound dye. An optimum dye concentration was found when Procion yellow HE-3G was bound to poly(ethylene glycol). When the same dye was bound to dextran, the number of dye molecules per dextran molecule influenced the effectiveness of the extraction.
D000110:Acetylcholinesterase / Q000302:isolation & purification; D000251:Adenosine Triphosphatases / Q000302:isolation & purification; D000818:Animals; D001923:Brain Chemistry; D002417:Cattle; D003377:Countercurrent Distribution; D003911:Dextrans; D011092:Polyethylene Glycols; D013385:Succinate Dehydrogenase / Q000302:isolation & purification; D013570:Synaptic Membranes / Q000032:analysis* / Q000201:enzymology; D013574:Synaptosomes / Q000032:analysis* / Q000201:enzymology; D014227:Triazines
D003911:Dextrans; D014227:Triazines; D011092:Polyethylene Glycols; C009794:procion yellow; D013385:Succinate Dehydrogenase; D000110:Acetylcholinesterase; D000251:Adenosine Triphosphatases