Open AccessPurification and properties of bovine liver seryl‐tRNA synthetase
Author(s) -
MIZUTANI Takaharu,
NARIHARA Takahiko,
HASHIMOTO Atsushi
Publication year - 1984
Publication title -
european journal of biochemistry
Language(s) - English
Resource type - Journals
eISSN - 1432-1033
pISSN - 0014-2956
DOI - 10.1111/j.1432-1033.1984.tb08331.x
Subject(s) - ammonium sulfate precipitation , affinity chromatography , sodium dodecyl sulfate , gel electrophoresis , chromatography , molecular mass , enzyme , column chromatography , biochemistry , chemistry , polyacrylamide gel electrophoresis , biology , size exclusion chromatography
Seryl‐tRNA synthetase was purified 1800‐fold from bovine liver extract by ultracentrifugation at 150000 × g , chromatography on DEAE‐cellulose, fractional precipitation with ammonium sulfate, gel chromatography on Sephacryl S‐300, adsorption chromatography on hydroxyapatite. affinity chromatography on blue‐Sepharose and finally on Mätrex gel red A. The relative molecular mass, M r , in the denatured state was estimated as 87000 by sodium dodecyl sulfate disc gel electrophoresis; in the active state the M r was estimated as 170000 for the dimeric native enzyme (α 2 type) by chromatography on Sephacryl S‐300. The amino acid composition of the enzyme was determined. The K m values for ATP and serine were 0.49 mM and 30 μM, respectively. The K m values for tRNA Ser IGA and tRNA Ser CmCA were 1.40 μM and 1.25 μM, respectively. Sequences common to the two isoaccepting tRNA Ser molecules are discussed in relation to the recognition mechanism of the purified seryl‐tRNA synthetase.