title: Purification of brain tubulin by affinity chromatography on immobilized lactoperoxidase.
abstract: Brain tubulin binds to lactoperoxidase coupled to Affigel 10 through a 10 A succinylated aminoalkyl spacer and can be eluted by an ionic strength gradient. The tubulin can be obtained about 90% electrophoretically pure in 2 to 3 h without glycerol or GTP. It retains its ability to bind colchicine. Compared to tubulin purified by the assembly-disassembly procedure, affinity-purified tubulin has a higher critical concentration for polymerization and the purified protein appears to be free of high molecular weight microtubule-associated proteins. Tubulin binding to the lactoperoxidase affinity column protects the colchicine binding site against decay at 4 degrees C, whereas the interaction of tubulin with soluble lactoperoxidase does not.
mesh_terms: D000818:Animals; D001923:Brain Chemistry; D002846:Chromatography, Affinity / Q000379:methods; D003078:Colchicine / Q000378:metabolism; D004229:Dithiothreitol; D004355:Drug Stability; D007700:Kinetics; D007784:Lactoperoxidase; D011485:Protein Binding; D051381:Rats; D014404:Tubulin / Q000302:isolation & purification* / Q000378:metabolism
chemical_list: D014404:Tubulin; D007784:Lactoperoxidase; D003078:Colchicine; D004229:Dithiothreitol