Catalog # EL7351

eEF2K Recombinant Protein

Lysate

Application / Dilution

WB	10 μl/lane

Size 100 μl

Eukaryotic elongation factor 2 (eEF2) catalyzes the translocation of peptidyl-tRNA from the A site to the P site on the ribosome. eEF2 kinase (eEF2K) phosphorylates and inactivates eEF2, resulting in the inhibition of peptide-chain elongation. eEF2K is normally dependent on Ca2+ ions and calmodulin, and can be activated by PKA in response to elevated cAMP levels during cell stress- or starvation-related conditions. Regulation of eEF2K occurs through phosphorylation at multiple sites. Ser-78 phosphorylation is required for calmodulin binding and eEF2K activity, while phosphorylation of Ser-500 is required for Ca2+/calmodulin-independent kinase activity. Thr-348 is an autophosphorylation site that is required for kinase activity. Inhibitory phosphorylation may also regulate eEF2K, since phosphorylation at Ser-359 by SAPK4/p38delta causes inactivation of eEF2K.

Human recombinant eEF2K is a full length protein (95 kDa) expressed in E. Coli. The protein is detected by rabbit polyclonal anti-eEF2K (C-terminus) antibody (Cat.#EP4661) and anti-eEF2K (Thr-348) (Cat.#EP4411).

References

Tavares, C.D. et al. (2012) Biochemistry. 51(11):2232.
Smith, E.M. & Proud, C.G. (2008) EMBO J. 27(7):1005.
Diggle, T.A., et al. (2001) Biochem J. 353(3):621.
Ryazanov, A.G. et al. (1997) Proc Natl Acad Sci. USA. 94:4884.

Recombinant human eEF2K untreated (lanes 1 & 4) and auto-phosphorylated in the presence of Ca2+ and Calmodulin (lanes 2 & 5). The eEF2K was dephosphorylated with lambda phosphatase (lanes 3 & 6). The blots were probed with anti-eEF2K (N-terminus) (lanes 1-3) or anti-eEF2K (Thr-348) (lanes 4-6). (Images provided by Dr. Kevin Dalby in the Dept. of Pharmacy at the University of Texas.)

eEF2K recombinant protein lysate is supplied at a concentration of 5 ng/µl in electrophoresis sample buffer (62.5 mM Tris pH 6.8, 2% SDS, 5% glycerol, 0.003% bromophenol blue, 0.9% β-mercaptoethanol). Store at –20°C. Do not boil or dilute. Stable for 1 year.

The products are are safely shipped at ambient temperature for both domestic and international shipments. Each product is guaranteed to match the specifications as indicated on the corresponding technical data sheet. Please store at -20C upon arrival for long term storage.

*All molecular weights (MW) are confirmed by comparison to Bio-Rad Rainbow Markers and to western blot mobilities of known proteins with similar MW.

This kit contains: