Eukaryotic initiation factor 5 from calf liver is a single polypeptide chain protein of M(r) = 62,000

P. Raychaudhuri, A. Chaudhuri, U. Maitra

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Abstract

Eukaryotic initiation factor 5 (eIF-5), which specifically catalyzes the joining of a 60 S ribosomal subunit to a 40 S initiation complex to form a functional 80 S initiation complex, has been purified from ribosomal salt wash proteins of calf liver. The purified factor exhibits only one polypeptide band of M(r) = 62,000 following electrophoresis in 10% polyacrylamide gels in the presence of sodium dodecyl sulfate. The native protein has a sedimentation coefficient of 4.2 S and a Stokes radius of 33 Å which is consistent with eIF-5 being a monomeric protein of M(r) = 58,000-62,000. Less pure preparations of eIF-5 elute in gel filtration columns with an apparent M(r) of 160,000-180,000 presumably due to association of eIF-5 with other high molecular weight proteins since eIF-5 activity present in such preparations can also be shown by gel electrophoretic separation under denaturing conditions to be associated with a 62,000-dalton protein. Furthermore, eIF-5 purified from calf liver extracts with or without a number of protease inhibitors is indistinguishable with regard to molecular weight and final specific activity of purified preparations. The purified factor catalyzes the hydrolysis of GTP present in 40 S initiation complexes in the absence of 60 S ribosomal subunits. The presence of 60 S ribosomal subunits neither stimulates nor inhibits the hydrolysis of GTP in the absence of Met-tRNA(f) or other components required for 40 S initiation complex formation. It can be calculated that 1 pmol of eIF-5 protein can catalyze the formation of at least 10 pmol of 80 S initiation complex under the conditions of in vitro initiation reactions.

Original languageEnglish (US)
Pages (from-to)2132-2139
Number of pages8
JournalJournal of Biological Chemistry
Volume260
Issue number4
StatePublished - 1985

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Eukaryotic Initiation Factor-5
Liver
Peptides
Ribosome Subunits
Proteins
Guanosine Triphosphate
Hydrolysis
Molecular Weight
Gels
Molecular weight
Liver Extracts
Protease Inhibitors
Electrophoresis
Sedimentation
Joining
Sodium Dodecyl Sulfate
Gel Chromatography
Salts
Association reactions

ASJC Scopus subject areas

  • Biochemistry

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Eukaryotic initiation factor 5 from calf liver is a single polypeptide chain protein of M(r) = 62,000. / Raychaudhuri, P.; Chaudhuri, A.; Maitra, U.

In: Journal of Biological Chemistry, Vol. 260, No. 4, 1985, p. 2132-2139.

Research output: Contribution to journalArticle

Raychaudhuri, P. ; Chaudhuri, A. ; Maitra, U. / Eukaryotic initiation factor 5 from calf liver is a single polypeptide chain protein of M(r) = 62,000. In: Journal of Biological Chemistry. 1985 ; Vol. 260, No. 4. pp. 2132-2139.
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abstract = "Eukaryotic initiation factor 5 (eIF-5), which specifically catalyzes the joining of a 60 S ribosomal subunit to a 40 S initiation complex to form a functional 80 S initiation complex, has been purified from ribosomal salt wash proteins of calf liver. The purified factor exhibits only one polypeptide band of M(r) = 62,000 following electrophoresis in 10{\%} polyacrylamide gels in the presence of sodium dodecyl sulfate. The native protein has a sedimentation coefficient of 4.2 S and a Stokes radius of 33 {\AA} which is consistent with eIF-5 being a monomeric protein of M(r) = 58,000-62,000. Less pure preparations of eIF-5 elute in gel filtration columns with an apparent M(r) of 160,000-180,000 presumably due to association of eIF-5 with other high molecular weight proteins since eIF-5 activity present in such preparations can also be shown by gel electrophoretic separation under denaturing conditions to be associated with a 62,000-dalton protein. Furthermore, eIF-5 purified from calf liver extracts with or without a number of protease inhibitors is indistinguishable with regard to molecular weight and final specific activity of purified preparations. The purified factor catalyzes the hydrolysis of GTP present in 40 S initiation complexes in the absence of 60 S ribosomal subunits. The presence of 60 S ribosomal subunits neither stimulates nor inhibits the hydrolysis of GTP in the absence of Met-tRNA(f) or other components required for 40 S initiation complex formation. It can be calculated that 1 pmol of eIF-5 protein can catalyze the formation of at least 10 pmol of 80 S initiation complex under the conditions of in vitro initiation reactions.",
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