Catalog # EP4411

eEF2K (Thr-348), phospho-specific Antibody

Rabbit Polyclonal

Application / Dilution

ELISA	1:2000
WB	1:1000

Size 100 μl

Species Reactivity Hu, Rt, Ms

MW 105 kDa

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/p38δ causes inactivation of eEF2K. Thus, multisite phospho-regulation of eEF2K may be important for proper control of eEF2K activity and protein translation.

References

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). In some lanes, the eEF2K was dephosphorylated with lambda phosphatase (lanes 3 & 6). The blots were probed with rabbit polyclonal anti-eEF2K (N-terminus) (lanes 1-3) or anti-eEF2K (Thr-348) (lanes 4-6). (Images provided by the laboratory of Dr. Kevin Dalby in the Dept. of Pharmacy at the University of Texas at Austin.)

Phospho-eEF2K (Thr-348) synthetic peptide (coupled to carrier protein) corresponds to amino acids surrounding Thr-348 in human eEF2K. This sequence is well conserved in rat and mouse eEF2K, and has low homology to other proteins.

*For more information, see UniProt Accession O00418

Rabbit polyclonal, affinity-purified antibody is supplied in 100µl phosphate-buffered saline, 50% glycerol, 1 mg/ml BSA, and 0.05% sodium azide. Store at –20°C. 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.

The antibody was affinity purified using phospho-eEF2K (Thr-348) peptide (without carrier). The antibody detects a 105 kDa* protein corresponding to the molecular mass of eEF2K on SDS-PAGE immunoblots of human recombinant eEF2K autophosphorylated in the presence of Ca2+ and Calmodulin. This reactivity is removed after lambda phosphatase treatment.

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

Product References:

Tavares, C.D. et al. (2017) J Biol Chem. 292(5):2032. (WB: human MDA-MB-231, eEF2K protein)
Fuchs, T. et al. (2017) Mol Psychiatry. 22(6):920. (WB: mouse hippocampus)
Tavares, C.D. et al. (2014) J Biol Chem. 289(34):23901. (WB: human breast cancer cells)
Tavares, C.D. et al. (2012) Biochemistry. 51(11):2232. (WB: human eEF2K activated, mutant)

This kit contains: