Cell Signaling Technology

Product Pathways - Translational Control

Phospho-eEF2k (Ser366) Antibody #3691

Applications Reactivity Sensitivity MW (kDa) Source
W IP H R Mk Endogenous 105 Rabbit

Applications Key:  W=Western Blotting  IP=Immunoprecipitation
Reactivity Key:  H=Human  R=Rat  Mk=Monkey
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-eEF2k (Ser366) Antibody detects endogenous levels of eEF2k only when phosphorylated at Ser366.

Source / Purification

Polyclonal antibodies are produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Ser366 of human eEF2k. Antibodies are purified by protein A and peptide affinity chromatography.

Western Blotting

Western Blotting

Western blot analysis of extracts from SW-13 cells (starved for 18 hours) treated with calf intestinal alkaline phosphatase (CIP) or 20% fetal bovine serum for 30 minutes, and extracts from HeLa cells (starved for 18 hours) treated with 200 nM TPA for 30 minutes, using Phospho-eEF2k (Ser366) Antibody (upper) or eEF2k Antibody #3692 (lower).

Background

Eukaryotic elongation factor 2 kinase (eEF2k) phosphorylates and inactivates eEF2, resulting in the inhibition of peptide-chain elongation (1). eEF2k is normally dependent on Ca2+ ions and calmodulin (2,3). It can be activated by PKA in response to elevated cAMP levels (4-6), which are generally increased in stress- or starvation-related conditions. eEF2k can also be regulated in response to a wide range of stimuli that promote cell growth and protein synthesis. This involves the phosphorylation of eEF2k by p90RSK and p70 S6 kinase at Ser366 or by SAPK4/p38delta at Ser359, leading to the inactivation of eEF2k (7,8), which facilitates the dephosphorylation of eEF2, and thus promotes translation.

  1. Ryazanov, A.G. et al. (1997) Proc. Natl. Acad. Sci. USA 94, 4884-4889.
  2. Nairn, A.C. et al. (1985) Proc. Natl. Acad. Sci. USA 82, 7839-7943.
  3. Palfrey, H.C. et al. (1987) J. Biol. Chem. 262, 9785-9792.
  4. Redpath, N.T. et al. (1993) Biochem. J. 293, 31-34.
  5. Diggle, T.A. et al. (1998) Biochem. J. 336, 525-529.
  6. Hovland, R. et al. (1999) FEBS Lett. 444, 97-101.
  7. Wang, X. et al. (2001) EMBO J. 20, 4370-4379.
  8. Knebel, A. et al. (2001) EMBO J. 20, 4360-4369.

Application References

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