Purification and characterization of a ribosome dissociation factor (eukaryotic initiation factor 6) from wheat germ.

D. W. Russell, L. L. Spremulli

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Abstract

A wheat germ ribosome dissociation factor, eukaryotic initiation factor 6 (eIF-6), has been purified almost to homogeneity from the 25 to 40% ammonium sulfate fraction of the postribosomal supernatant. This dissociation factor is distinct from initiation factor eIF-3 and its chromatographic properties permit its separation from the known wheat germ initiation factors. Under certain conditions, eIF-6 stimulates the incorporation of amino acids into polypeptides in a partially fractionated wheat germ cell-free system. The eight-step purification procedure developed includes chromatography on DEAE-cellulose, phosphocellulose, Sephadex G-75, and hydroxyapatite and yields a dissociation factor more than 80% pure. The purified factor is composed of a single polypeptide chain with a molecular weight of approximately 23,000 as determined by gel filtration chromatography and by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. It is an acidic protein which is heat labile and is inactivated by treatment with N-ethylmaleimide. The dissociation factor is much more effective in preventing the reassociation of 40 S and 60 S ribosomal subunits than in directly dissociating 80 S ribosomes. Like Escherichia coli IF-3, about 10 pmol of the dissociation factor are required to dissociate 1 pmol of ribosomes.

Original languageEnglish (US)
Pages (from-to)8796-8800
Number of pages5
JournalJournal of Biological Chemistry
Volume254
Issue number18
StatePublished - Sep 25 1979

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Ribosomes
Triticum
Purification
Peptide Initiation Factors
Ribosome Subunits
DEAE-Cellulose Chromatography
Peptides
Chromatography
Ethylmaleimide
Cell-Free System
Ammonium Sulfate
Durapatite
Germ Cells
Sodium Dodecyl Sulfate
Gel Chromatography
Polyacrylamide Gel Electrophoresis
Hot Temperature
Molecular Weight
DEAE-Cellulose
Escherichia coli

ASJC Scopus subject areas

  • Biochemistry

Cite this

Purification and characterization of a ribosome dissociation factor (eukaryotic initiation factor 6) from wheat germ. / Russell, D. W.; Spremulli, L. L.

In: Journal of Biological Chemistry, Vol. 254, No. 18, 25.09.1979, p. 8796-8800.

Research output: Contribution to journalArticle

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