Affinity Labeling of the GDP/GTP Binding Site in Thermus Thermophilus Elongation Factor Tu

  • Marcus E. Peter
  • Mathias Sprinzl


Affinity labeling of T. thermophilus EF-Tu•GDP(GTP) complexes by in situ oxidation with periodate results in specific modification of lysine residues 52, 137 and 325. Residue 52 of the native EF-Tu is modified by both GDPOXI and GTPOXI., but it cannot be modified by GDPOXI if the protein is cleaved at arginine 59. Residue 137 is preferentially modified by GTPOXI and is essentially unreactive toward GDPOXI Cleavage of EF-Tu at position 59 renders an amino acid residue be very sensitive to reaction with GDPOXI which is essentially unreactive in the native protein. Residue 325 is a minor reaction site accessible from both GDPOXI and GTPOXI in the native protein. EF-Tu nicked at position 59 does not show any reaction with GDPOXI at lysine 325. Photoirradiation leads to crosslinking of the guanine moiety with region 181–190 of T. thermophilus EF-Tu. When the above affinity labeling results are fitted into the available three dimensional models of E. coli EF-Tu•GDP and H-ras protooncogen product p21•GDP complexes the following points emerge:
  1. 1)

    There is a high homology in the folding of the structural domains in both proteins.

  2. 2)

    The loop region connecting helix A with the β-sheet b (La Cour et al., 1985) of elongation factor Tu is placed in the vicinity of the bound GDP/GTP and in analogy to p21 it probably forms a part of a binding pocket for the nucleotide. This loop corresponds to the “effector loop” of G-proteins.

  3. 3)

    Cleavage at position 59 of EF-Tu leads to a conformational change resulting in altered reactivity of GDPOXI. towards lysine residues adjacent to the nucleotide binding pocket.



Lysine Residue Elongation Factor Nucleotide Binding Site Cyanogen Bromide Additional Amino Acid 
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Copyright information

© Springer Science+Business Media New York 1989

Authors and Affiliations

  • Marcus E. Peter
    • 1
  • Mathias Sprinzl
    • 1
  1. 1.Laboratorium für Biochemie und Bayreuther Institut für makromolekulare ForschungUniversität BayreuthBayreuthGermany

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