Biological characterization of various forms of elongation factor 1 from rabbit reticulocytes.
Two forms of elongation factor 1 (EF-1) have been tested for a variety of biological functions. One form, EF-1H, is a high-molecular-weight aggregate (Mr greater than 500,000) containing four distinct polypeptides (alpha, beta, gamma, delta). The other form, EF-1 alpha, consists of a single polypeptide which is the same as the alpha subunit of EF-1H. Both EF-1 alpha and EF-1H function catalytically in binding Phe-tRNA to ribosomes, and in poly(U)-directed polyphenylalanine synthesis. The activity of EF-1 alpha is enhanced in polyphenylalanine synthesis by a complementary component, EF-1 beta delta. It is also shown that EF-1 beta delta can facilitate an exchange of EF-1 alpha-bound GDP for GTP. The EF-1 alpha dissociation constants for GDP and GTP were 0.47 and 0.55 microM respectively, while the EF-1H dissociation constants for GDP and GTP were 2.0 and 1.6 microM, respectively. Thus, while EF-1 alpha and EF-1H had approximately the same affinities for GDP and GTP, the EF-1 alpha dissociation constants were about fourfold lower than the EF-1H dissociation constants. Attempts to isolate complexes of EF-1 alpha or EF-1H with GTP and Phe-tRNA or with GTP, Phe-tRNA, and ribosomes were unsuccessful using either Millipore filters, gel filtration, or sucrose density gradients. The results presented in this report, along with studies from other laboratories, strengthen the hypothesis that the general mechanism of the elongation cycle is similar in eucaryotes and procaryotes.
D000818:Animals; D002499:Centrifugation, Density Gradient; D006153:Guanosine Diphosphate / Q000378:metabolism; D006160:Guanosine Triphosphate / Q000378:metabolism; D046911:Macromolecular Substances; D008970:Molecular Weight; D010452:Peptide Biosynthesis; D020648:Peptide Elongation Factor 1; D010445:Peptide Elongation Factors / Q000097:blood / Q000502:physiology*; D010455:Peptides; D011072:Poly U / Q000378:metabolism; D012346:RNA, Transfer, Amino Acyl / Q000378:metabolism; D011817:Rabbits; D012156:Reticulocytes / Q000032:analysis*; D012270:Ribosomes / Q000378:metabolism
D046911:Macromolecular Substances; D020648:Peptide Elongation Factor 1; D010445:Peptide Elongation Factors; D010455:Peptides; D012346:RNA, Transfer, Amino Acyl; D006153:Guanosine Diphosphate; D011072:Poly U; C012001:polyphenylalanine; D006160:Guanosine Triphosphate