The
guanine nucleotide exchange factor,
elongation factor 1 beta gamma (
EF-1 beta gamma) has been purified from Artemia
cysts using an improved method. The
protein consists of two distinct
polypeptides with relative molecular masses of 26,000 (EF-1 beta) and 46,000 (EF-1 gamma). A
nucleoside diphosphate phosphotransferase activity often found in
EF-1 beta gamma preparations has been completely separated from the actual
guanine nucleotide exchange stimulatory activity of
EF-1 beta gamma, thus indicating that
nucleotide diphosphate phosphotransferase is not an intrinsic property of
EF-1 beta. Both
EF-1 beta gamma and
EF-1 beta have been shown to stimulate the following three reactions to a comparable degree: (a) exchange of
GDP bound to
EF-1 alpha with exogenous
GDP; (b)
EF-1 alpha-dependent binding of Phe-
tRNA to ribosomes; (c)
poly(U)-dependent poly(
phenylalanine) synthesis. However, a significantly higher
nucleotide exchange rate was observed in the presence of
EF-1 beta gamma compared to
EF-1 beta alone. Concerning
elongation factor 1 gamma (EF-1 gamma) the following observations were made. In contrast to
EF-1 beta, pure
EF-1 gamma is rather insoluble in aqueous
buffers, but the tendency to precipitate can be partially suppressed by the addition of
detergents. In particular,
EF-1 gamma partitions solely into the
detergent phase of
Triton X-114 solutions.
EF-1 gamma is also more susceptible to spontaneous, specific fragmentation. It is remarkably that about 5% of the cellular pool of
EF-1 beta gamma was found to be present in membrane fractions, under conditions where no
EF-1 alpha was detectable in these fractions. Furthermore it was noted that
EF-1 beta gamma copurified strongly with
tubulin on
DEAE-cellulose. Moreover, it was observed that from a mixture of
EF-1 beta gamma and
tubulin,
EF-1 gamma coprecipitates with
tubulin using a non-denaturating immunoprecipitation technique. These findings suggest that
EF-1 gamma has a hydrophobic domain and interacts with membrane and cytoskeleton structures in the cell.