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The addition or removal of phosphate groups from proteins is the most common chemical modification that regulates cellular processes. These modifications can affect the structure, activity, stability, and localization of proteins within cells as well as their interactions with other proteins.
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Membrane lipids such as phosphatidylinositol (PI) are precursors for several membrane-bound and soluble second messengers. Specific kinases phosphorylate PI and produce phosphorylated inositol phospholipids. One such inositol phospholipids are the  phosphatidylinositol-4,5 bisphosphate [PI(4,5)P2], present in the inner half of the lipid bilayer. Upon ligand binding, GPCR stimulates Gq proteins to turn on phospholipase Cꞵ. Activated phospholipase Cꞵ cleaves PI(4,5)P2 and...
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Heterotrimeric G proteins are guanine nucleotide-binding proteins. As the name suggests, heterotrimeric G proteins are composed of three subunits: alpha, beta, and gamma. They remain GDP-bound or GTP-bound inside the cells and switch between inactive/active states. The Gα subunit possesses the nucleotide-binding pocket that binds guanine nucleotides and switches between GDP or GTP-bound states. In contrast, the Gꞵ and Gγ subunits are always bound together with high...
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Proteins undergo chemical modifications that trigger changes in the charge, structure, and conformation of the proteins. Phosphorylation, acetylation, glycosylation, nitrosylation, ubiquitination, lipidation, methylation, and proteolysis are various protein modifications that regulate protein activity. Such modifications are usually enzyme-driven.
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EF-G Activation by Phosphate Analogs.

Enea Salsi1, Elie Farah1, Dmitri N Ermolenko1

  • 1Department of Biochemistry and Biophysics & Center for RNA Biology, School of Medicine and Dentistry, University of Rochester, Rochester, NY 14642, USA.

Journal of Molecular Biology
|April 12, 2016
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Summary

Elongation factor G (EF-G) binding to the ribosome requires GTP. GTP hydrolysis, not directly coupled to translocation, triggers EF-G release, facilitating protein synthesis.

Keywords:
Beryllium fluorideEF-GGTP hydrolysisRibosomemRNA translocation

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Area of Science:

  • Molecular Biology
  • Protein Synthesis
  • Biochemistry

Background:

  • Elongation factor G (EF-G) is crucial for ribosome function.
  • EF-G facilitates tRNA and mRNA translocation during translation.
  • The role of EF-G GTP hydrolysis in translocation is debated.

Purpose of the Study:

  • To investigate the role of GTP hydrolysis in EF-G mediated translocation.
  • To determine the mechanism by which EF-G interacts with the ribosome.

Main Methods:

  • Utilized phosphate group analogs (BeF3-, AlF4-) to mimic transition states.
  • Compared translocation rates with GTP and non-hydrolyzable GTP analogs.
  • Studied EF-G•GDP binding to the ribosome.

Main Results:

  • EF-G•GDP alone does not stably bind or induce translocation.
  • EF-G•GDP with BeF3- or AlF4- promotes tRNA/mRNA translocation.
  • mRNA translocation rates are similar with GTP and GDP•BeF3-.

Conclusions:

  • GTP binding, not hydrolysis, induces the translocation-competent state of EF-G.
  • GTP hydrolysis likely triggers EF-G release from the ribosome.
  • Protein translocation mechanism clarified: GTP binding activates, hydrolysis releases.