Valyl-tRNA synthetase from rabbit liver. II. The enzyme derived from the high-Mr complex displays hydrophobic as well as polyanion-binding properties.
The preceding paper (Bec, G., Kerjan, P., Zha, X.D., and Waller, J.P. (1989) J. Biol. Chem. 264, 21131-21137) described the purification to apparent homogeneity from rabbit liver, of a heterotypic complex comprising valyl-tRNA synthetase and Elongation Factor 1H. In the present study, valyl-tRNA synthetase was dissociated and separated from the other components of this complex by hydroxylapatite chromatography in the presence of 0.5 M NaSCN. The properties of the homogeneous mammalian enzyme were compared to those of the corresponding enzyme from yeast. Both behaved as monomeric entities, with apparent molecular masses of 140 and 125 kDa, respectively. Furthermore, both displayed strong affinity toward the polyanionic support heparin-Ultrogel, a property not manifested by the corresponding prokaryotic enzyme. However, unlike the yeast enzyme, that of mammalian origin additionally exhibited hydrophobic properties, as reflected by its affinity toward phenyl-Sepharose. A structural model is proposed according to which mammalian valyl-tRNA synthetase has conserved the polycationic N-terminal domain that distinguishes the corresponding lower eukaryotic enzyme from its prokaryotic counterpart, while acquiring a hydrophobic domain most likely responsible for its association to Elongation Factor 1H.
Bec
G
G
Laboratoire d'Enzymologie, Centre National de la Recherche Scientifique, Gif-sur-Yvette, France.
Waller
J P
JP
D000604:Amino Acyl-tRNA Synthetases / Q000378:metabolism*; D000818:Animals; D002499:Centrifugation, Density Gradient; D002845:Chromatography; D002846:Chromatography, Affinity; D002850:Chromatography, Gel; D017886:Durapatite; D004591:Electrophoresis, Polyacrylamide Gel; D005990:Glycerol; D006882:Hydroxyapatites; D008099:Liver / Q000201:enzymology*; D008970:Molecular Weight; D020648:Peptide Elongation Factor 1; D010445:Peptide Elongation Factors / Q000302:isolation & purification / Q000378:metabolism*; D011817:Rabbits; D012261:Ribonucleoproteins / Q000378:metabolism*; D014637:Valine-tRNA Ligase / Q000302:isolation & purification / Q000378:metabolism*
D006882:Hydroxyapatites; D020648:Peptide Elongation Factor 1; D010445:Peptide Elongation Factors; D012261:Ribonucleoproteins; D017886:Durapatite; D000604:Amino Acyl-tRNA Synthetases; D014637:Valine-tRNA Ligase; D005990:Glycerol