Purified eukaryotic initiation factor 2 from calf liver consists of two polypeptide chains of 48,000 and 38,000 daltons.

Stringer, Evan A.; Chaudhuri, Asok; Maitra, Umadas

doi:10.1016/s0021-9258(18)50248-9

Cited by 56 publications

(2 citation statements)

References 23 publications

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“…The translation initiation factor eIF2γ was purified from 12 day old chick embryos ribosome as described by Stringer et al (12). The crude 0.4 M KCl ribosome extract was fractionated by (NH 4 ) 2 SO 4 , dialyzed and chromatographed on a DEAE-sepharose column (12). Mismatch-specific thymine-DNA glycosylase was purified from HeLa cells as described by Neddermann and Jiricny (13).…”

Section: Purification Of Other Proteinsmentioning

confidence: 99%

Demethylation of DNA by purified chick embryo 5-methylcytosine-DNA glycosylase requires both protein and RNA

Frémont

Siegmann

Gaulis

et al. 1997

Nucleic Acids Research

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We have previously purified and characterized a 5-methylcytosine (5-MeC)-DNA glycosylase from 12 day old chick embryos [Jost,J.P. et al. (1995) J. Biol. Chem. 270, 9734-9739]. The activity of the purified enzyme is abolished upon treatment with proteinase K and ribonuclease A. RNA copurifies with 5-MeC-DNA glycosylase activity throughout all chromatographic steps and preparative gel electrophoresis. RNA with a length of approximately 300-500 nucleotides was isolated from the gel purified enzyme. Upon extensive treatment with proteinase K, the gel eluted and labeled RNA did not show any significant change in molecular mass. The purified RNA incubated alone or in the presence of Mg2+and deoxyribonucleotide phosphates had no 5-MeC-DNA glycosylase or demethylating activities. However, activity of 5-MeC-DNA glycosylase could be restored when the purified RNA was incubated with the inactive protein, free of RNA.

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Section: Purification Of Other Proteinsmentioning

confidence: 99%

Demethylation of DNA by purified chick embryo 5-methylcytosine-DNA glycosylase requires both protein and RNA

Frémont

Siegmann

Gaulis

et al. 1997

Nucleic Acids Research

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show abstract

“…Other Methods. eIF-2 activity was measured by the ability of the factor to promote GTP-dependent binding of [35S]-Met-tRNAf to a nitrocellulose membrane filter as described previously (Stringer et al, 1979). Protein was determined by the method of Bradford (1976) using reagents obtained from Bio-Rad Laboratories.…”

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confidence: 99%

Purification and Characterization of Bacterially Expressed Mammalian Translation Initiation Factor 5 (eIF-5): Demonstration That eIF-5 Forms a Specific Complex with eIF-2

Chaudhuri¹,

Das²,

Maitra³

1994

Biochemistry

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Eukaryotic translation initiation factor 5, eIF-5, has been purified to apparent electrophoretic homogeneity from overproducing Escherichia coli cells expressing the cDNA of the initiation factor under the control of the T7 promoter-T7 RNA polymerase system. Purified recombinant eIF-5 mimics natural eIF-5 isolated from mammalian cells in size, in specific activity, in its ability to catalyze the hydrolysis of GTP bound to the 40S initiation complex, and in the subsequent joining with 60S ribosomal subunits to form the 80S initiation complex. Further characterization of eIF-5 demonstrates that eIF-5 specifically associates with eIF-2, forming an eIF-2.eIF-5 complex. The protein complex sediments in glycerol gradients with an apparent M(r) of 160,000, suggesting that the two proteins associate in a 1:1 stoichiometry. The association between the two initiation factors is highly specific. Addition of 32P-labeled eIF-5 to a partially purified rabbit reticulocyte initiation factor preparation that contained, in addition to eIF-2 and eIF-5, other initiation factors and many other proteins resulted in the specific binding of labeled eIF-5 only to eIF-2, forming a 160-kDa protein complex. In agreement with these observations, we found that in crude initiation factor preparations derived from rabbit reticulocyte lysates, eIF-5 was present as an eIF-2.eIF-5 complex. The significance of eIF-2.eIF-5 complex formation in the overall mechanism of GTP hydrolysis in protein synthesis initiation is discussed.

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Roles of Eukaryotic Initiation Factor 2 and Eukaryotic Initiation Factor 2 Ancillary Protein Factors in Eukaryotic Protein Synthesis Initiation

Gupta¹,

Ahmad²,

Chakrabarti³

et al. 1987

Translational Regulation of Gene Expression

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Purified eukaryotic initiation factor 2 from calf liver consists of two polypeptide chains of 48,000 and 38,000 daltons.

Cited by 56 publications

References 23 publications

Demethylation of DNA by purified chick embryo 5-methylcytosine-DNA glycosylase requires both protein and RNA

Demethylation of DNA by purified chick embryo 5-methylcytosine-DNA glycosylase requires both protein and RNA

Purification and Characterization of Bacterially Expressed Mammalian Translation Initiation Factor 5 (eIF-5): Demonstration That eIF-5 Forms a Specific Complex with eIF-2

Roles of Eukaryotic Initiation Factor 2 and Eukaryotic Initiation Factor 2 Ancillary Protein Factors in Eukaryotic Protein Synthesis Initiation

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