Purification and characterization of the Saccharomyces cerevisiae mitochondrial leucyl-tRNA synthetase.
We have purified the product of the NAM2 gene, the mitochondrial leucyl-tRNA synthetase, from yeast mitochondria. The purified protein cross-reacts with antibodies raised against the product of a LacZ/NAM2 gene fusion and antibodies raised against the purified Escherichia coli leucyl-tRNA synthetase. The mass as determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis is about 100 kDa, consistent with the size predicted by the gene sequence (102 kDa). The N-terminal sequence of the protein has been determined and shows that the first nine amino acids predicted by the gene sequence have been removed, probably during transport into the mitochondria.
Zagorski
W
W
Centre de Génétique Moléculaire, Centre National de la Recherche Scientifique associé a Université Pierre et Maire Curie, Gif-sur-Yvette, France.
Castaing
B
B
Herbert
C J
CJ
Labouesse
M
M
Martin
R
R
Slonimski
P P
PP
D000255:Adenosine Triphosphate / Q000378:metabolism; D000595:Amino Acid Sequence; D015153:Blotting, Western; D002845:Chromatography; D003429:Cross Reactions; D004591:Electrophoresis, Polyacrylamide Gel; D005800:Genes, Fungal; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D007930:Leucine / Q000378:metabolism; D007935:Leucine-tRNA Ligase / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification / Q000378:metabolism*; D008928:Mitochondria / Q000201:enzymology*; D008969:Molecular Sequence Data; D012441:Saccharomyces cerevisiae / Q000201:enzymology* / Q000235:genetics; D013696:Temperature
D000255:Adenosine Triphosphate; D007935:Leucine-tRNA Ligase; D007930:Leucine