Aminoacylation of tRNA-Leu species from Escherichia coli and from the cytoplasm, chloroplasts and mitochondria of Phaseolus vulgaris by homologous and heterologous enzymes.
Leucyl-tRNA synthetase from Phaseolus vulgaris chloroplasts could be separated from its cytoplasmic counterpart upon chromatography on hydroxyapatite, but the cytoplasmic and mitochondrial leucyl-tRNA synthetases could not be distinguished. The tRNALeu species from the various plant cell compartments and from Escherichia coli were aminoacylated using either homologous or heterologous enzymes; the levels of aminoacylation and the profiles of the leucyl-tRNAs upon reverse-phase chromatography were studied. Cytoplasmic tRNALeu species could be aminoacylated by the cytoplasmic or by the mitochondrial enzymes and in both cases yielded two peaks upon reverse-phase chromatography (RPC-5). But they could not be charged by the chloroplast-specific or by the E. coli enzynes. Mitochondrial tRNALeu species could be charged by the mitochondrial or by the cytoplasmic enzymes and in both cases yielded four peaks upon reverse phase (RPC-5) chromatography. But they could not be aminoacylated using the chloroplast-specific or the E. coli leucyl-tRNA synthetases. Chloroplastic tRNALeu species can be divided into two classes: the first class contains four isoacceptor species which can be charged by the cytoplasmic or mitochondrial enzymes, but not by the chloroplast-specific or the E. coli enzymes; the second class contains three chloroplast-specific tRNALeu species which can be charged by the chloroplast-specific or the E. coli enzymes but not by the cytoplasmic or the mitochondrial enzymes. There are five isoacceptor tRNALeu species in E. coli; all are charged by the E. coli or the chloroplast-specific enzymes, while only one is aminoacylated by the plant cytoplasmic or mitochondrial enzymes.
D000604:Amino Acyl-tRNA Synthetases / Q000302:isolation & purification / Q000378:metabolism*; D002499:Centrifugation, Density Gradient; D002736:Chloroplasts / Q000201:enzymology*; D002845:Chromatography; D002852:Chromatography, Ion Exchange; D003593:Cytoplasm / Q000201:enzymology; D004926:Escherichia coli / Q000201:enzymology*; D007700:Kinetics; D007930:Leucine; D008274:Magnesium / Q000494:pharmacology; D008928:Mitochondria / Q000201:enzymology*; D010944:Plants / Q000201:enzymology*
D000604:Amino Acyl-tRNA Synthetases; D007930:Leucine; D008274:Magnesium