Cell Signaling Technology

Product Pathways - Translational Control

Phospho-eEF2 (Thr56) Antibody #2331

Applications Reactivity Sensitivity MW (kDa) Source
W H M R Hm Mk C Endogenous 95 Rabbit

Applications Key:  W=Western Blotting
Reactivity Key:  H=Human  M=Mouse  R=Rat  Hm=Hamster  Mk=Monkey  C=Chicken
Species cross-reactivity is determined by western blot. Species enclosed in parentheses are predicted to react based on 100% sequence homology.

Protocols

Specificity / Sensitivity

Phospho-eEF2 (Thr56) Antibody detects endogenous levels of eEF2 only when phosphorylated at Thr56. It does not recognize eEF2 phosphorylated at other sites.

Source / Purification

Polyclonal antibodies are produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Thr56 of human eEF2. Antibodies are purified by protein A and peptide affinity chromatography.

Western Blotting

Western Blotting

Western blot analysis of extracts from C6 cells, untreated or forskolin-treated (10 µM for 60 minutes), using Phospho-eEF2 (Thr56) Antibody (upper) or eEF2 Antibody #2332 (lower).

Background

Eukaryotic elongation factor 2 (eEF2) catalyzes the translocation of peptidyl-tRNA from the A site to the P site on the ribosome. It has been shown that phosphorylation of eEF2 at threonine 56 by eEF2 kinase inhibits its activity (1-4). eEF2 kinase is normally dependent on Ca2+ ions and calmodulin (5,6). eEF2 kinase can also be activated by PKA in response to elevated cAMP levels (7-9), which are generally increased in stress- or starvation-related conditions. A variety of treatments known to raise intracellular Ca2+ or cAMP levels have been shown to result in increased phosphorylation of eEF2, and thus to inhibit peptide-chain elongation. The inactive phosphorylated eEF2 can be converted to its active nonphosphorylated form by a protein phosphatase, most likely a form of protein phosphatase-2A (PP-2A). Insulin, which activates protein synthesis in a wide range of cell types, induces rapid dephosphorylation of eEF2 through mTOR signaling and may involve modulation of the activity of the PP-2A or the eEF2 kinase or both (10).

  1. Nairn, A.C. and Palfrey, H.C. (1987) J. Biol. Chem. 262, 17299-17303.
  2. Ryazanov, A.G. et al. (1988) Nature 334, 170-173.
  3. Carlberg, U. et al. (1990) Eur. J. Biochem. 191, 639-645.
  4. Redpath, N.T. et al. (1993) Eur. J. Biochem. 213, 689-699.
  5. Nairn, A.C. et al. (1985) Proc. Natl. Acad. Sci. USA 82, 7939-7943.
  6. Palfrey, H.C. et al. (1987) J. Biol. Chem. 262, 9785-9792.
  7. Redpath, N.T. and Proud, C.G. (1993) Biochem. J. 293, 31-34.
  8. Diggle, T. et al. (1998) Biochem. J. 336, 525-529.
  9. Hovland, R. et al. (1999) FEBS Lett. 444, 97-101.
  10. Proud, C. (2000) Translational Control of Gene Expression. Cold Spring Harbor Laboratory Press, NY, 719-739.

Application References

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For Research Use Only. Not For Use In Diagnostic Procedures.

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