Isolation of prolyl-tRNA synthetase as a free form and as a form associated with glutamyl-tRNA synthetase

Shu Mei Ting, P. Bogner, John David Dignam

Research output: Contribution to journalArticle

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Abstract

Rat liver prolyl-tRNA synthetase was purified as a dimer of Mr 60,000 subunits not associated with other aminoacyl-tRNA synthetases and as a form associated with glutamyl-tRNA synthetase. Proteolysis of the dimeric enzyme generated a less active form with Mr 52,000 subunits and an inactive form with Mr 40,000 subunits. A second species was isolated with polypeptides of Mr 60,000 and 150,000. This form dissociated during gel filtration chromatography being partially resolved into the Mr 150,000 and 60,000 components; glutamyl-tRNA synthetase was associated with the larger polypeptide and prolyl-tRNA synthetase with the smaller component. Antibodies against the Mr 60,000 polypeptide reacted with the Mr 60,000 and 150,000 polypeptides. Gel filtration of extracts revealed multiple forms of prolyl- and glutamyl-tRNA synthetase. Antibody against the Mr 60,000 component detected the Mr 60,000 and 150,000 polypeptides throughout the chromatogram; these forms could be partially separated by polyethylene glycol fractionation. The Mr 150,000 and 60,000 polypeptides were detected by Western blot analysis of crude extracts prepared under several conditions. Antibody to prolyl-tRNA synthetase reacted with a Mr 150,000 polypeptide of the aminoacyl-tRNA synthetase core complex identified previously as glutamyl-tRNA synthetase.

Original languageEnglish
Pages (from-to)17701-17709
Number of pages9
JournalJournal of Biological Chemistry
Volume267
Issue number25
Publication statusPublished - Sep 5 1992

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Glutamate-tRNA Ligase
Peptides
Amino Acyl-tRNA Synthetases
Gel Chromatography
Antibodies
Gels
Proteolysis
Fractionation
Chromatography
prolyl T RNA synthetase
Complex Mixtures
Dimers
Liver
Rats
Western Blotting

ASJC Scopus subject areas

  • Biochemistry

Cite this

Isolation of prolyl-tRNA synthetase as a free form and as a form associated with glutamyl-tRNA synthetase. / Ting, Shu Mei; Bogner, P.; Dignam, John David.

In: Journal of Biological Chemistry, Vol. 267, No. 25, 05.09.1992, p. 17701-17709.

Research output: Contribution to journalArticle

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