[Phenylalanyl-tRNA synthetase of wheat embryos. Purification, molecular weight, structure, properties].
From wheat germ, a phenylalanyl-tRNA synthetase (E.C.6.1.1.20) has been isolated and purified 187 fold by means of ammonium sulfate fractionation (40-50 per cent) followed by Sephadex G-200 gel filtration, chromatographies on DEAE-cellulose and hydroxyapatite. The enzyme appears to be homogeneous on Sephadex G-200 molecular filtration and polyacrylamide gel electrophoresis. Molecular weight determinations by sucrose gradient centrifugation, gel filtration and gel electrophoresis give an average of 250 00 daltons. The enzyme is dissociated in 1 per cent sodium dodecyl sulfate into two different equimolar components of 80 000 and 50 000 daltons ; this result suggests that the phenylalanyl-tRNA synthetase has a subunit structure : alpha2 beta2. Dissociation with sodium dodecyl sulfate and dithiothreitol gives four other components, probably resulting from the breakdown of the subunits. Optima values of pH, Mg2+ and K+ concentrations, effect of SH-compnents, kinetic parameters have been determined in the aminoacylation reaction. Physical and catalytic properties of wheat germ phenylalanyl-tRNA synthetase appear very similar to those of the yeast and E. coli enzymes.
D000604:Amino Acyl-tRNA Synthetases / Q000302:isolation & purification*; D002850:Chromatography, Gel; D004591:Electrophoresis, Polyacrylamide Gel; D007700:Kinetics; D046911:Macromolecular Substances; D008274:Magnesium; D008970:Molecular Weight; D010455:Peptides / Q000032:analysis; D010652:Phenylalanine-tRNA Ligase / Q000302:isolation & purification* / Q000378:metabolism; D011188:Potassium; D011487:Protein Conformation; D012639:Seeds / Q000201:enzymology; D013439:Sulfhydryl Reagents / Q000494:pharmacology; D014908:Triticum / Q000201:enzymology*; D014461:Ultracentrifugation
D046911:Macromolecular Substances; D010455:Peptides; D013439:Sulfhydryl Reagents; D000604:Amino Acyl-tRNA Synthetases; D010652:Phenylalanine-tRNA Ligase; D008274:Magnesium; D011188:Potassium
Phénylalanyl-trna synthétase des embryons de blé. Purification, masse molaire, structure, propriétés