Separation of Myosin Subfragment-1 into Fractions Containing g1 Chain and g3 Chain by Sepharose-Adipic Acid Hydrazide-ATP Column Chromatography
- 1 May 1976
- journal article
- research article
- Published by Oxford University Press (OUP) in The Journal of Biochemistry
- Vol. 79 (5) , 1049-1052
- https://doi.org/10.1093/oxfordjournals.jbchem.a131146
Abstract
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 KC1. Ninety-nine per cent of subfragment-1 was adsorbed on the column in this medium. Forty-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 KC1. The former fraction contained g3 chain and the latter g1 chain. These fractions were apparently the same as the two 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 105 g of protein. The preparation of the two subfragment-1 fractions is described.Keywords
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