Properties and substrate specificities of the phenylalanyl-transfer-ribonucleic acid synthetases of Aesculus species.
1. Phenylalanyl-tRNA synthetases have been partially purified from cotyledons of seeds of Aesculus californica, which contains 2-amino-4-methylhex-4-enoic acid, and from four other species of Aesculus that do not contain this amino acid. The A. californica preparation was free from other aminoacyl-tRNA synthetases, and the contaminating synthetase activity in preparations from A. hippocastanum was decreased to acceptable limits by conducting assays of pyrophosphate exchange activity in 0.5m-potassium chloride. 2. The phenylalanyl-tRNA synthetase from each species activated 2-amino-4-methylhex-4-enoic acid with K(m) 30-40 times that for phenylalanine. The maximum velocity for 2-amino-4-methylhex-4-enoic acid was only 30% of that for phenylalanine with the A. californica enzyme, but the maximum velocities for the two substrates were identical for the other four species. 3. 2-Amino-4-methylhex-4-enoic acid was not found in the protein of A. californica, so discrimination against this amino acid probably occurs in the step of transfer to tRNA, though subcellular localization, or subsequent steps of protein synthesis could be involved. 4. Crotylglycine, methallylglycine, ethallylglycine, 2-aminohex-4,5-dienoic acid, 2-amino-5-methylhex-4-enoic acid, 2-amino-4-methylhex-4-enoic acid, beta-(thien-2-yl)alanine, beta-(pyrazol-1-yl)alanine, phenylserine and m-fluorophenylalanine were substrates for pyrophosphate exchange catalysed by the phenylalanyl-tRNA synthetases of A. californica or A. hippocastanum. Allylglycine, phenylglycine and 2-amino-4-phenylbutyric acid were inactive.
D000409:Alanine / Q000378:metabolism; D002122:Calcium Chloride / Q000494:pharmacology; D002208:Caproates / Q000378:metabolism; D002848:Chromatography, DEAE-Cellulose; D005998:Glycine / Q000378:metabolism; D007700:Kinetics; D008025:Ligases / Q000032:analysis*; D008274:Magnesium / Q000494:pharmacology; D010649:Phenylalanine / Q000378:metabolism*; D010944:Plants / Q000201:enzymology*; D011188:Potassium / Q000494:pharmacology; D012343:RNA, Transfer / Q000378:metabolism*; D012639:Seeds / Q000201:enzymology; D012694:Serine / Q000378:metabolism; D013045:Species Specificity
D002208:Caproates; D012694:Serine; D010649:Phenylalanine; D012343:RNA, Transfer; D008025:Ligases; D008274:Magnesium; D002122:Calcium Chloride; D000409:Alanine; D011188:Potassium; D005998:Glycine