E. coli elongation factor Tu bound to a GTP analogue displays an open conformation equivalent to the GDP-bound form.

Publisher: Oxford University Press Country of Publication: England NLM ID: 0411011 Publication Model: Print Cited Medium: Internet ISSN: 1362-4962 (Electronic) Linking ISSN: 03051048 NLM ISO Abbreviation: Nucleic Acids Res Subsets: MEDLINE

Publication: 1992- : Oxford : Oxford University Press
Original Publication: London, Information Retrieval ltd.

Protein Conformation*; Escherichia coli/*chemistry ; Guanosine Triphosphate/*chemistry ; Peptide Elongation Factor Tu/*chemistry; Crystallography, X-Ray ; Escherichia coli/genetics ; GTP Phosphohydrolases/chemistry ; GTP Phosphohydrolases/genetics ; Guanosine Diphosphate/chemistry ; Guanosine Triphosphate/analogs & derivatives ; Peptide Elongation Factor Tu/genetics ; Protein Biosynthesis/genetics ; RNA, Messenger/chemistry ; RNA, Messenger/genetics ; Ribosomes/chemistry ; Ribosomes/genetics

According to the traditional view, GTPases act as molecular switches, which cycle between distinct 'on' and 'off' conformations bound to GTP and GDP, respectively. Translation elongation factor EF-Tu is a GTPase essential for prokaryotic protein synthesis. In its GTP-bound form, EF-Tu delivers aminoacylated tRNAs to the ribosome as a ternary complex. GTP hydrolysis is thought to cause the release of EF-Tu from aminoacyl-tRNA and the ribosome due to a dramatic conformational change following Pi release. Here, the crystal structure of Escherichia coli EF-Tu in complex with a non-hydrolysable GTP analogue (GDPNP) has been determined. Remarkably, the overall conformation of EF-Tu·GDPNP displays the classical, open GDP-bound conformation. This is in accordance with an emerging view that the identity of the bound guanine nucleotide is not 'locking' the GTPase in a fixed conformation. Using a single-molecule approach, the conformational dynamics of various ligand-bound forms of EF-Tu were probed in solution by fluorescence resonance energy transfer. The results suggest that EF-Tu, free in solution, may sample a wider set of conformations than the structurally well-defined GTP- and GDP-forms known from previous X-ray crystallographic studies. Only upon binding, as a ternary complex, to the mRNA-programmed ribosome, is the well-known, closed GTP-bound conformation, observed.

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R35 GM118139 United States GM NIGMS NIH HHS; R01 GM080376 United States GM NIGMS NIH HHS

0 (RNA, Messenger)
146-91-8 (Guanosine Diphosphate)
86-01-1 (Guanosine Triphosphate)
EC 3.6.1.- (GTP Phosphohydrolases)
EC 3.6.1.- (Peptide Elongation Factor Tu)

Date Created: 20180815 Date Completed: 20190628 Latest Revision: 20190628

20221213

PMC6144822

10.1093/nar/gky697

30107565