Site-Directed Mutagenesis of Elongation Factor Tu
Elongation factor Tu (EF-Tu), a monomeric protein of 393 amino acid residues (M.W. 43,000), is the most abundant protein in E.coli and one of the best studied guanine nucleotide binding proteins, a family of enzymes involved in signal transduction in higher and lower organisms (for references see Bosch et al., 1984; Gilman, 1984; Parmeggiani and Swart, 1985; Bourne, 1986). These proteins bind GTP and GDP, are able to hydrolyze GTP and show typical homologies in their primary structures, especially in the N-terminal 150–200 amino acids. The finding that the ras p21 protein, a mutant variant of which is responsible for oncogenic transformation, is a guanine nucleotide binding protein (Scolnick et al., 1979) further emphasizes the importance of this family. EF-Tu is an essential component of protein biosynthesis, acting as the carrier of aa-tRNA to the ribosome (for references, see Milled: & Weissbach, 1977). As with the other guanine nucleotide binding proteins, GTP induces the active form of the factor: only EF-Tu·GTP is capable of interacting with aa-tRNA, forming a ternary complex.
KeywordsHydrolysis Polypeptide Phenylalanine Guanine Phospho
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- Bosch, L., Kraal B., Van der Meide, P.H., Duisterwinkel, F.J., Van Noort, 1983, Prog. Nucleic Acid Res. Mol. Biol., 20; 91–126.Google Scholar
- Fasano, O., Bruns, W., Crechet, J.B., Sander, G., Parmeggiani, A., 1978, Eur. J. Biochem., 124; 53–58.Google Scholar
- Halliday, K.R., 1983, J. Cyclic Nucl. Phosph. Res. 9; 435–448.Google Scholar
- Lebermann, R., Egner, U. 1984, Embo J., 3; 339–341.Google Scholar
- Master, S.B., Stroud, R.M., Bourne, H.R. 1986, Protein Engineering, 1; 47–54.Google Scholar
- Miller, D.L., Weissbach H. 1977, in: “Molecular Mechanisms in Protein Biosynthesis,” Weissbach, H., Pestka, S., eds., pp. 323–373, Academic Press, New York.Google Scholar
- Parmeggiani, A., Anborgh, P.H., Canceill, D., Jacquet, J., Jonak, J., Merola, M., Mortensen, K.K., Swart, G.W.M. 1986, in “Structure, Function and Genetics of Ribosomes,” Hardesty, B. & Kramer, G., eds., pp. 672–685, Springer Verlag, New York.Google Scholar
- Permeggiani, A., Swart, G.W.M., Mortensen, K.K., Jensen, M., Dente, L., Cortese, R., 1987, Proc. Natl. Acad. Sci., in print, U.S.A.Google Scholar
- Swart, G.W.M., Merola, M., Guesnet, J., Parmeggiani, A. 1984, in “Metabolism and Enzymology of Nucleic Acids Including Gene Manipulations 5, Zelinka, J. & Balan, J., eds., pp. 277–288, Slovak Academy of Sciences, Bratislava.Google Scholar
- Swart, G.W.M., 1987, in “The Polypeptide Chain Elongation Factor Tu from E.coli. Characterization of Mutants and Protein Engineering,” Ph.D. Thesis, Leiden, pp. 83–119.Google Scholar