Conformational change of elongation factor Tu (EF-Tu) induced by antibiotic binding. Crystal structure of the complex between EF-Tu.GDP and aurodox.
Aurodox is a member of the family of kirromycin antibiotics, which inhibit protein biosynthesis by binding to elongation factor Tu (EF-Tu). We have determined the crystal structure of the 1:1:1 complex of Thermus thermophilus EF-Tu with GDP and aurodox to 2.0-A resolution. During its catalytic cycle, EF-Tu adopts two strikingly different conformations depending on the nucleotide bound: the GDP form and the GTP form. In the present structure, a GTP complex-like conformation of EF-Tu is observed, although GDP is bound to the nucleotide-binding site. This is consistent with previous proposals that aurodox fixes EF-Tu on the ribosome by locking it in its GTP form. Binding of EF-Tu.GDP to aminoacyl-tRNA and mutually exclusive binding of kirromycin and elongation factor Ts to EF-Tu can be explained on the basis of the structure. For many previously observed mutations that provide resistance to kirromycin, it can now be understood how they prevent interaction with the antibiotic. An unexpected feature of the structure is the reorientation of the His-85 side chain toward the nucleotide-binding site. We propose that this residue stabilizes the transition state of GTP hydrolysis, explaining the acceleration of the reaction by kirromycin-type antibiotics.
Vogeley
L
L
Department of Structural Biology and Crystallography, Institute of Molecular Biotechnology, Beutenbergstrasse 11, D-07745 Jena, Germany.
Palm
G J
GJ
Mesters
J R
JR
Hilgenfeld
R
R
D000900:Anti-Bacterial Agents / Q000737:chemistry / Q000378:metabolism*; D000899:Aurodox / Q000737:chemistry* / Q000378:metabolism*; D001665:Binding Sites; D006153:Guanosine Diphosphate / Q000378:metabolism*; D006160:Guanosine Triphosphate / Q000378:metabolism; D006165:Guanylyl Imidodiphosphate / Q000378:metabolism; D007930:Leucine; D008958:Models, Molecular; D008968:Molecular Conformation; D010444:Peptide Elongation Factor Tu / Q000737:chemistry* / Q000378:metabolism*; D011485:Protein Binding; D011487:Protein Conformation; D011994:Recombinant Proteins / Q000378:metabolism; D016963:Thermus thermophilus / Q000378:metabolism; D014443:Tyrosine
D000900:Anti-Bacterial Agents; D011994:Recombinant Proteins; D000899:Aurodox; D006153:Guanosine Diphosphate; D006165:Guanylyl Imidodiphosphate; D014443:Tyrosine; D006160:Guanosine Triphosphate; D010444:Peptide Elongation Factor Tu; D007930:Leucine