The effect of F-actin on the binding and hydrolysis of guanine nucleotide by Dictyostelium elongation factor 1A

Brian T. Edmonds, Andrea Bell, Jeffrey Wyckoff, John S. Condeelis, Thomas S. Leyh

Research output: Contribution to journalArticle

36 Citations (Scopus)

Abstract

Indirect evidence implicates actin as a cofactor in eukaryotic protein synthesis. The present study directly examines the effects of F-actin on the biochemical properties of eukaryotic elongation factor 1A (eEF1A, formerly EF1α), a major actin-binding protein. The basal mechanism of eEF1A alone is determined under physiological conditions with the critical finding that glycerol and guanine nucleotide are required to prevent protein aggregation and loss of enzymatic activity. The dissociation constants (K(d)) for GDP and GTP are 2.5 μM and 0.6 μM, respectively, and the k(cat) of GTP hydrolysis is 1.0 x 10-3 s-1. When eEF1A binds to F-actin, there is a 7-fold decrease in the affinity for guanine nucleotide and an increase of 35% in the rate of GTP hydrolysis. Based upon our results and the relevant cellular concentrations, the predominant form of cellular eEF1A is calculated to be GTP·eEF1A·F-actin. We conclude that F-actin does not significantly modulate the basal enzymatic properties of eEF1A; however, actin may still influence protein synthesis by sequestering GTP·eEF1A away from interactions with its known translational ligands, e.g. aminoacyl-tRNA and ribosomes.

Original languageEnglish (US)
Pages (from-to)10288-10295
Number of pages8
JournalJournal of Biological Chemistry
Volume273
Issue number17
DOIs
StatePublished - Apr 24 1998

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Peptide Elongation Factors
Dictyostelium
Guanine Nucleotides
Actins
Hydrolysis
Guanosine Triphosphate
Microfilament Proteins
Proteins
Transfer RNA
Ribosomes
Glycerol
Agglomeration
Ligands

ASJC Scopus subject areas

  • Biochemistry

Cite this

The effect of F-actin on the binding and hydrolysis of guanine nucleotide by Dictyostelium elongation factor 1A. / Edmonds, Brian T.; Bell, Andrea; Wyckoff, Jeffrey; Condeelis, John S.; Leyh, Thomas S.

In: Journal of Biological Chemistry, Vol. 273, No. 17, 24.04.1998, p. 10288-10295.

Research output: Contribution to journalArticle

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