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

被引:40
作者
Edmonds, BT
Bell, A
Wyckoff, J
Condeelis, J
Leyh, TS
机构
[1] Yeshiva Univ Albert Einstein Coll Med, Dept Anat & Struct Biol, Bronx, NY 10461 USA
[2] Yeshiva Univ Albert Einstein Coll Med, Dept Biochem, Bronx, NY 10461 USA
关键词
D O I
10.1074/jbc.273.17.10288
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
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 EF1 alpha), 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 mu M and 0.6 mu M, 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.
引用
收藏
页码:10288 / 10295
页数:8
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