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

doi:10.1016/0006-291X(86)90372-4

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

Chemistry and Biochemistry

Research output: Contribution to journal › Article › peer-review

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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-tRNA_i, 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 language	English
Pages (from-to)	1160-1166
Number of pages	7
Journal	Biochemical and Biophysical Research Communications
Volume	134
Issue number	3
DOIs	https://doi.org/10.1016/0006-291X(86)90372-4
State	Published - 13 Feb 1986

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

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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AU - Siekierka, John

AU - Ochoa, Severo

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AB - 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

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