Science Advances

Supplementary Materials

This PDF file includes:

  • table S1. Interplay between EF-Tu, pEF-TuT382, and guanosine nucleotides.
  • table S2. Parameters obtained from the stopped-flow kinetic measurements.
  • table S3. X-ray data collection and refinement statistics.
  • table S4. SAXS parameters of the different species.
  • table S5. Further parameters obtained after PDA analysis of the experimental FRET data.
  • table S6. Relevant input parameters for the spFRET simulations.
  • table S7. Simulated spFRET parameters obtained after the dynamic PDA analysis.
  • table S8. Solvent accessibility of experimentally validated phosphorylation sites in the E. coli proteome.
  • table S9. The oligonucleotides used for the construction of the EF-Tu mutants EF-TuT382E, EF-TuT61E, EF-TuT382E/S222C, and EF-TuT61E/S222C.
  • fig. S1. In vitro phosphorylation of EF-Tu by Doc.
  • fig. S2. ITC titrations of EF-Tu and phosphorylated EF-Tu with nucleotides, EF-Ts and Glu-tRNAGlu.
  • fig. S3. Stopped-flow kinetics of the EF-Tu and phosphorylated EF-Tu interaction with nucleotides.
  • fig. S4. Interaction of EF-Tu and phosphorylated EF-Tu with aa-tRNAs.
  • fig. S5. Structural effects of phosphorylation of EF-Tu at Thr382.
  • fig. S6. Characterization of the labeling of EF-Tu with ATTO 488 and Alexa Fluor 647.
  • fig. S7. Multiparameter graphs obtained after spFRET analysis of EF-Tu and phosphorylated EF-Tu in the presence of GDP and GDPNP.
  • fig. S8. Multiparameter graphs obtained after spFRET analysis of the EF-Tu phosphomimetic mutants in the presence of GDP and GDPNP.
  • fig. S9. Static versus dynamic PDA analysis of the EF-Tu phosphomimetic mutants in the presence of GDP and GDPNP.
  • fig. S10. X-ray structure of the EF-Tu phosphomimetic mutants.
  • fig. S11. Analysis of the conservation of the phosphorylation sites across the EF-Tu superfamily.

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