Purification and properties of the guanine nucleotide exchange factor (GEF) from HeLa cells and its role in the initiation of protein synthesis

Thomas M. Mariano, John Siekierka, Severo Ochoa

Research output: Contribution to journalArticleResearchpeer-review

9 Citations (Scopus)

Abstract

A guanine nucleotide exchange factor (GEF)1 1 The abbreviations used are: eIF-2, eukaryotic initiation factor 2; GEF, GDP exchange factor; HCI, heme-controlled translational inhibitor (an eIF-2α kinase); DAI, double-stranded (ds) RNA-activated translational inhibitor (another eIF-2α kinase); Met-tRNAi, eukaryotic initiator methionyl-tRNA; SDS-PAGE, sodium dodecylsulfate-polyacrylamide gel electrophoresis., catalyzing the exchange of GDP bound to initiation factor eIF-2 for GTP, has been isolated from S3 HeLa cells as the eIF-2·GEF complex and extensively purified by procedures originally developed for purification of GEF from rabbit reticulocytes. The HeLa cell factor resembles rabbit reticulocyte eIF-2·GEF in polypeptide composition, catalytic activity, and inactivation by α-phosphorylated eIF-2.

Original languageEnglish
Pages (from-to)1160-1166
Number of pages7
JournalBiochemical and Biophysical Research Communications
Volume134
Issue number3
DOIs
StatePublished - 13 Feb 1986

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Guanine Nucleotide Exchange Factors
eIF-2 Kinase
HeLa Cells
Purification
Reticulocytes
Polyacrylamide Gel Electrophoresis
Proteins
Prokaryotic Initiation Factor-2
Eukaryotic Initiation Factor-2
RNA, Transfer, Met
Rabbits
Peptide Initiation Factors
Double-Stranded RNA
Human computer interaction
Transfer RNA
Guanosine Triphosphate
Electrophoresis
Heme
Catalyst activity
Sodium

Cite this

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abstract = "A guanine nucleotide exchange factor (GEF)1 1 The abbreviations used are: eIF-2, eukaryotic initiation factor 2; GEF, GDP exchange factor; HCI, heme-controlled translational inhibitor (an eIF-2α kinase); DAI, double-stranded (ds) RNA-activated translational inhibitor (another eIF-2α kinase); Met-tRNAi, eukaryotic initiator methionyl-tRNA; SDS-PAGE, sodium dodecylsulfate-polyacrylamide gel electrophoresis., catalyzing the exchange of GDP bound to initiation factor eIF-2 for GTP, has been isolated from S3 HeLa cells as the eIF-2·GEF complex and extensively purified by procedures originally developed for purification of GEF from rabbit reticulocytes. The HeLa cell factor resembles rabbit reticulocyte eIF-2·GEF in polypeptide composition, catalytic activity, and inactivation by α-phosphorylated eIF-2.",
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Purification and properties of the guanine nucleotide exchange factor (GEF) from HeLa cells and its role in the initiation of protein synthesis. / Mariano, Thomas M.; Siekierka, John; Ochoa, Severo.

In: Biochemical and Biophysical Research Communications, Vol. 134, No. 3, 13.02.1986, p. 1160-1166.

Research output: Contribution to journalArticleResearchpeer-review

TY - JOUR

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