Separation of myosin subfragment-1 into fractions containing g1 chani and g2 chain by Sepharose-adipic acid hydrazide-ATP column chromatography.
Subfragment-1 prepared by chymotryptic digestion of myosin was applied to a column of Sepharose-adipic acid hydrazide-ATP in 1 mM EDTA, 10 mM Tris-HCL (PH 7.6), and 40 mM KCL. Ninety-nine per cent of subfragment-1 was adsorbed on the column in this medium. Fourty-three per cent of the applied protein was eluted with 6 mM ADP in the above buffer and then 52% was eluted with 1 mM EDTA, 10 mM Tris-HCL (pH 7.6), AND 0.7 M KCL. The former fraction contained g3 chain and the latter g1 chain. These fractions were apparently the same as the components, p2 and p1, respectively, isolated by ion-exchange chromatography using DEAE-cellulose (Yagi & Otani (1974) J. Biochem. 76, 365-373). No significant difference of ADP binding was found between the two fractions, both could bind about 0.5 mole per 10(5) g of protein. The preparation of the two subfragment-1 fractions is described.
D000244:Adenosine Diphosphate; D000272:Adipates; D000818:Animals; D001665:Binding Sites; D002846:Chromatography, Affinity; D004591:Electrophoresis, Polyacrylamide Gel; D009132:Muscles; D009218:Myosins / Q000302:isolation & purification*; D011485:Protein Binding; D011817:Rabbits
D000272:Adipates; D000244:Adenosine Diphosphate; D009218:Myosins