Open AccessArginyl‐tRNA Synthetase from Baker's Yeast. Purification and Some Properties
Author(s) -
GANGLOFF Jean,
SCHUTZ Annie,
DIRHEIMER Guy
Publication year - 1976
Publication title -
european journal of biochemistry
Language(s) - English
Resource type - Journals
eISSN - 1432-1033
pISSN - 0014-2956
DOI - 10.1111/j.1432-1033.1976.tb10403.x
Subject(s) - aminoacylation , transfer rna , enzyme , yeast , biochemistry , polyacrylamide gel electrophoresis , saccharomyces cerevisiae , chemistry , arginine , sodium , gel electrophoresis , chromatography , amino acid , substrate (aquarium) , electrophoresis , rna , biology , organic chemistry , ecology , gene
Arginyl‐tRNA synthetase from baker's yeast (Saccharomyces cerevisiae, strain 836) was obtained pure by a large‐scale preparative method, which involves four chromatographic columns and one preparative polyacrylamide gel electrophoretic step. The enzyme has a high specific activity (9000 U/ mg) and consists of a single polypeptide chain of molecular weight approximately 73000 as determined by polyacrylamide gel electrophoresis in the presence of sodium dodecylsulphate. Amino acid analysis of the enzyme permitted calculation of the absorption coefficient of arginyl‐tRNA synthetase (A 1 mg /ml 280 nm = 1.26). Concerning kinetic parameters of the enzyme we found the following K m values: 0.28 μM, 300 μM, 1.5 μM for tRNA Arg 111 , ATP and arginine in the aminoacylation reaction, and 1400 μM, 2.5 μM, and 50 μM for ATP, arginine and PP i in the ATP‐PP i exchange reaction. Arginyl‐ tRNA synthetase requires tRNA Arg 111 to catalyse the ATP‐PP i exchange reaction.