Rapid purification of threonyl-tRNA synthetase from Saccharomyces carlsbergensis by affinity elution from phosphocellulose. 1978

H Yamada

Threonyl-tRNA synthetase [EC 6.1.1.3] of Saccharomyces carlsbergensis was highly purified by a simple enzyme-substrate affinity method. After calcium-gel treatment of the crude enzyme extract, ammonium sulfate precipitation, and removal of polynucleotides with DEAE-cellulose, the enzyme was nonspecifically adsorbed onto phosphocellulose (P-cellulose), and then eluted specifically with a linear concentration gradient of threonine tRNA from torula yeast (Candida (Torulopsis) utilis). The enzyme was purified 303-fold from the calcium-gel supernatant. This purification method is very simple and time-saving. The purified enzyme gave a single band on SDS-gel electrophoresis, indicating a molecular weight of 82,000, and exhibited a molecular weight of about 150,000 by gel filtration. This suggests that the enzyme consists of two identical subunits. Some kinetic properties of the pure enzyme are described.

UI MeSH Term Description Entries
D007700 Kinetics The rate dynamics in chemical or physical systems.
D008970 Molecular Weight The sum of the weight of all the atoms in a molecule. Molecular Weights,Weight, Molecular,Weights, Molecular
D002846 Chromatography, Affinity A chromatographic technique that utilizes the ability of biological molecules, often ANTIBODIES, to bind to certain ligands specifically and reversibly. It is used in protein biochemistry. (McGraw-Hill Dictionary of Scientific and Technical Terms, 4th ed) Chromatography, Bioaffinity,Immunochromatography,Affinity Chromatography,Bioaffinity Chromatography
D000604 Amino Acyl-tRNA Synthetases A subclass of enzymes that aminoacylate AMINO ACID-SPECIFIC TRANSFER RNA with their corresponding AMINO ACIDS. Amino Acyl T RNA Synthetases,Amino Acyl-tRNA Ligases,Aminoacyl Transfer RNA Synthetase,Aminoacyl-tRNA Synthetase,Transfer RNA Synthetase,tRNA Synthetase,Acyl-tRNA Ligases, Amino,Acyl-tRNA Synthetases, Amino,Amino Acyl tRNA Ligases,Amino Acyl tRNA Synthetases,Aminoacyl tRNA Synthetase,Ligases, Amino Acyl-tRNA,RNA Synthetase, Transfer,Synthetase, Aminoacyl-tRNA,Synthetase, Transfer RNA,Synthetase, tRNA,Synthetases, Amino Acyl-tRNA
D012440 Saccharomyces A genus of ascomycetous fungi of the family Saccharomycetaceae, order SACCHAROMYCETALES. Saccharomyce
D013056 Spectrophotometry, Ultraviolet Determination of the spectra of ultraviolet absorption by specific molecules in gases or liquids, for example Cl2, SO2, NO2, CS2, ozone, mercury vapor, and various unsaturated compounds. (McGraw-Hill Dictionary of Scientific and Technical Terms, 4th ed) Ultraviolet Spectrophotometry
D013914 Threonine-tRNA Ligase An enzyme that activates threonine with its specific transfer RNA. EC 6.1.1.3. Threonyl T RNA Synthetase,Thr-tRNA Ligase,Threonyl-tRNA Synthetase,Ligase, Thr-tRNA,Ligase, Threonine-tRNA,Synthetase, Threonyl-tRNA,Thr tRNA Ligase,Threonine tRNA Ligase,Threonyl tRNA Synthetase

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