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PMID: 2263451 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Interaction of the isolated domain II/III of Thermus thermophilus elongation factor Tu with the nucleotide exchange factor EF-Ts.

Nucleic acids research ·Vol. 18 ·No. 23 ·1990-12-11 ·Pages 6889-93

Peter ME, Reiser CO, Schirmer NK, Kiefhaber T, Ott G, Grillenbeck NW, Sprinzl M

Abstract

The middle and C-terminal domain (domain II/III) of elongation factor Tu from Thermus thermophilus lacking the GTP/GDP binding domain have been prepared by treating nucleotide-free protein with Staphylococcus aureus V8 protease. The isolated domain II/III of EF-Tu has a compact structure and high resistance against tryptic treatment and thermal denaturation. As demonstrated by circular dichroism spectroscopy, the isolated domain II/III does not contain any alpha-helical structure. Nucleotide exchange factor, EF-Ts, was found to interact with domain II/III, whereas the binding of aminoacyl-tRNA, GDP and GTP to this EF-Tu fragment could not be detected.

MeSH Terms
Circular Dichroism Guanosine Diphosphate/metabolism Guanosine Triphosphate/metabolism Nucleic Acid Denaturation Peptide Elongation Factor Tu/chemistry,isolation & purification,metabolism Peptide Elongation Factors/metabolism RNA, Transfer, Amino Acyl/metabolism Serine Endopeptidases/metabolism Temperature Thermus/metabolism Trypsin/metabolism X-Ray Diffraction
Chemicals
Peptide Elongation Factors RNA, Transfer, Amino Acyl elongation factor Ts Guanosine Diphosphate Guanosine Triphosphate Serine Endopeptidases glutamyl endopeptidase Trypsin Peptide Elongation Factor Tu
Authors & Affiliations
7 authors, click to expand affiliations / ORCID
Peter M E
Laboratorium für Biochemie, Universität Bayreuth, FRG.
Reiser C O
Schirmer N K
Kiefhaber T
Ott G
Grillenbeck N W
Sprinzl M
References (34)
34 references, click to expand
  1. Cleavage of structural proteins during the assembly of the head of bacteriophage T4.
    Nature. 1970 Aug 15;227(5259):680-5 PMID: 5432063
  2. Structural determination of the functional sites of E. coli elongation factor Tu.
    Biochim Biophys Acta. 1990 Aug 27;1050(1-3):203-8 PMID: 2207145
  3. Spectrophotometric determination of protein concentration in cell extracts containing tRNA's and rRNA's.
    Anal Biochem. 1973 Aug;54(2):454-63 PMID: 4737496
  4. Conformational alteration of protein synthesis elongation factor EF-Tu by EF-Ts and by kirromycin.
    Proc Natl Acad Sci U S A. 1977 Aug;74(8):3264-7 PMID: 269389
  5. Studies on polypeptide-chain-elongation factors from an extreme thermophile, Thermus thermophilus HB8. 1. Purification and some properties of the purified factors.
    Eur J Biochem. 1978 Dec;92(2):509-19 PMID: 738277
  6. Studies on polypeptide-chain-elongation factors from an extreme thermophile, Thermus thermophilus HB8. 3. Molecular properties.
    Eur J Biochem. 1978 Dec;92(2):533-43 PMID: 33049
  7. Enzymic binding of aminoacyl-tRNA to Escherichia coli ribosomes using modified tRNA species and tRNA fragments.
    Methods Enzymol. 1979;60:615-28 PMID: 379537
  8. Aminoacyl transfer ribonucleic acid binding site of the bacterial elongation factor Tu.
    Biochemistry. 1980 May 13;19(10):2108-12 PMID: 6990972
  9. A computer-assisted model for estimating protein secondary structure from circular dichroic spectra: comparison of animal lactate dehydrogenases.
    Anal Biochem. 1980 May 1;104(1):160-7 PMID: 7386876
  10. Determination of protein secondary structure in solution by vacuum ultraviolet circular dichroism.
    J Mol Biol. 1980 Apr;138(2):149-78 PMID: 7411608
  11. Identification of a histidine residue near the aminoacyl transfer ribonucleic acid binding site of elongation factor Tu.
    Biochemistry. 1981 Aug 4;20(16):4663-6 PMID: 6913406
  12. The accuracy of protein biosynthesis is limited by its speed: high fidelity selection by ribosomes of aminoacyl-tRNA ternary complexes containing GTP[gamma S].
    Proc Natl Acad Sci U S A. 1982 Aug;79(16):4922-6 PMID: 6750613
  13. N-Tosyl-L-phenylalanylchloromethane reacts with cysteine 81 in the molecule of elongation factor Tu from Escherichia coli.
    FEBS Lett. 1982 Dec 27;150(2):485-8 PMID: 6761150
  14. Bacterial elongation factor Ts: isolation and reactivity with elongation factor Tu.
    J Bacteriol. 1983 Mar;153(3):1266-71 PMID: 6681813
  15. Specific alterations of the EF-Tu polypeptide chain considered in the light of its three-dimensional structure.
    EMBO J. 1984 Jan;3(1):113-20 PMID: 6323160
  16. Relative affinities of all Escherichia coli aminoacyl-tRNAs for elongation factor Tu-GTP.
    J Biol Chem. 1984 Apr 25;259(8):5010-6 PMID: 6370998
  17. A kinetic analysis of the interaction of elongation factor Tu with guanosine nucleotides and elongation factor Ts.
    J Biol Chem. 1984 Nov 10;259(21):12997-3003 PMID: 6386807
  18. Direct determination of the association constant between elongation factor Tu X GTP and aminoacyl-tRNA using fluorescence.
    Biochemistry. 1985 Jan 29;24(3):692-700 PMID: 3888260
  19. A second tRNA binding site on elongation factor Tu is induced while the factor is bound to the ribosome.
    Proc Natl Acad Sci U S A. 1985 May;82(10):3212-6 PMID: 3923474
  20. Structure of the GDP domain of EF-Tu and location of the amino acids homologous to ras oncogene proteins.
    Science. 1985 Oct 4;230(4721):32-6 PMID: 3898365
  21. Properties of a genetically engineered G domain of elongation factor Tu.
    Proc Natl Acad Sci U S A. 1987 May;84(10):3141-5 PMID: 3554231
  22. A structural model for the alpha-subunit of transducin. Implications of its role as a molecular switch in the visual signal transduction mechanism.
    FEBS Lett. 1987 Aug 10;220(1):15-22 PMID: 3038611
  23. G proteins: transducers of receptor-generated signals.
    Annu Rev Biochem. 1987;56:615-49 PMID: 3113327
  24. Sequence and identification of the nucleotide binding site for the elongation factor Tu from Thermus thermophilus HB8.
    Nucleic Acids Res. 1987 Nov 25;15(22):9263-77 PMID: 3317278
  25. Molecular cloning and sequence determination of the tuf gene coding for the elongation factor Tu of Thermus thermophilus HB8.
    Eur J Biochem. 1987 Dec 30;170(1-2):93-8 PMID: 2826164
  26. Three-dimensional structure of an oncogene protein: catalytic domain of human c-H-ras p21.
    Science. 1988 Feb 19;239(4842):888-93 PMID: 2448879
  27. Crosslinking of elongation factor Tu to tRNA(Phe) by trans-diamminedichloroplatinum (II). Characterization of two crosslinking sites on EF-Tu.
    FEBS Lett. 1989 Mar 13;245(1-2):194-200 PMID: 2494073
  28. Affinity labeling of the GDP/GTP binding site in Thermus thermophilus elongation factor Tu.
    Biochemistry. 1988 Dec 27;27(26):9132-9 PMID: 3242617
  29. Structure of the guanine-nucleotide-binding domain of the Ha-ras oncogene product p21 in the triphosphate conformation.
    Nature. 1989 Sep 21;341(6239):209-14 PMID: 2476675
  30. Interaction of elongation factor Tu from Escherichia coli with aminoacyl-tRNA carrying a fluorescent reporter group on the 3' terminus.
    Eur J Biochem. 1989 Sep 15;184(2):345-52 PMID: 2676533
  31. Identification of the N-tosyl-L-phenylalanyl chloromethylketone modification site in Thermus thermophilus elongation factor Tu.
    FEBS Lett. 1989 Nov 6;257(2):219-22 PMID: 2583265
  32. Structure of E. coli glutaminyl-tRNA synthetase complexed with tRNA(Gln) and ATP at 2.8 A resolution.
    Science. 1989 Dec 1;246(4934):1135-42 PMID: 2479982
  33. Mapping the effector region in Thermus thermophilus elongation factor Tu.
    Biochemistry. 1990 Mar 20;29(11):2876-84 PMID: 2189498
  34. Studies on polypeptide elongation factors from Escherichia coli. II. Purification of factors Tu-guanosine diphosphate, Ts, and Tu-Ts, and crystallization of Tu-guanosine diphosphate and Tu-Ts.
    J Biol Chem. 1972 Nov 10;247(21):7029-37 PMID: 4563073
Article Info
Journal
Nucleic acids research
Abbr.
Nucleic Acids Res
ISSN
0305-1048
DOI
10.1093/nar/18.23.6889
Published
1990-12-11
Pages
6889-93
Language
English
Region
England
NLM ID
0411011
PMCID
PMC332746
Subset
IM
External Links
PubMed source DOI: 10.1093/nar/18.23.6889 PMC Full Text View journal details
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