Catalog # EK6910


eEF2K Phospho-Regulation

Antibody Sampler Kit

$395

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 and Ser-445 are two of the major autophosphorylation sites required for kinase activity. Inhibitory phosphorylation may also regulate eEF2K, since phosphorylation at Ser-359 by SAPK4/p38δ causes inactivation of eEF2K. In addition, Ser-398 phosphorylation is required for eEFK2 induced autophagy. Thus, multisite phospho-regulation of eEF2K may be important for proper control of eEF2K activity and protein translation.

References
Diggle, T.A., et al. (2001) Biochem J. 353(3):621.
Smith, E.M. & Proud, C.G. (2008) EMBO J. 27(7):1005.
Tavares, C.D. et al. (2012) Biochemistry. 51(11):2232.
Cheng, Y. et al. (2013) Autophagy. 9(2):208.
Rabbit polyclonal antibodies are supplied in phosphate-buffered saline (PBS), 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.



*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:

EP4451 Kameshima, S et al.(2015)Am J Phys Hea Cir Phys.308(10):1298 WB: rat lung tissue
EP4411 Tavares, C.D. et al. (2012) Biochemistry. 51(11):2232. WB: human eEF2K activated, mutant
EP4451 Tavares, C.D. et al. (2012) Biochemistry. 51(11):2232. WB: human eEF2K activated, mutant

Western blot image of human Jurkat cells treated with calyculin A (100 nM) for 30 min. (lanes 1-6), then some lanes of the blot were treated with lambda phosphatase (lanes 2, 4 & 6). The blot was probed with rabbit polyclonals anti-eEF2K (C-terminus) (lanes 1 & 2), anti-eEF2K (Ser-445) phospho-specific (lanes 3 & 4), and anti-eEF2K (Ser-398) phospho-specific (lanes 5 & 6).

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.)



This kit contains:

CATALOG#DESCRIPTIONSIZEAPPLICATIONSSPECIES REACTIVITYMW (kDa)
eEF2K (Ser-78), phospho-specific Rabbit pAb50 μlWB, EHu, Rt, Ms105
eEF2K (Thr-348), phospho-specific Rabbit pAb 50 μlWB, EHu, Rt, Ms105
eEF2K (Ser-359), phospho-specific Rabbit pAb 50 μlWB, EHu, Rt, Ms105
eEF2K(Ser-398), phospho-specific Rabbit pAb50 μlWB, EHu, Rt, Ms105
eEF2K (Ser-445), phospho-specific Rabbit pAb50 μlWB, EHu, Rt, Ms, F105
eEF2K (Ser-500), phospho-specific Rabbit pAb50 μlWB, EHu, Rt, Ms105
KIT SUMMARY

The eEF2K phospho-regulation kit can be used to detect phosphorylation of Ser-78, Ser-359, Thr-348, Ser-398, Ser-445, and Ser-500 in eEF2K. Also, anti-eEF2K (C-terminus) rabbit polyclonal (Cat.# EP4661) is available separately for comparison of total eEF2K expression relative to site-specific phosphorylation level.