eef2k inhibitor  (MedChemExpress)

Journal: Molecular cell

Article Title: PQBP1 promotes translational elongation and regulates hippocampal mGluR-LTD by suppressing eEF2 phosphorylation.

doi: 10.1016/j.molcel.2021.01.032

Figure Lengend Snippet: Figure 4. PQBP1 suppresses eEF2K-mediated eEF2 phosphorylation (A) In vitro eEF2 phosphorylation assay showing that PQBP1, but not PQBP1D45-81, suppresses eEF2 phosphorylation. Top: western blots of the p-eEF2 level. Center: Coomassie blue staining for purified MBP-eEF2, GST-eEF2K, and PQBP1 proteins. Bottom: quantification of the relative level of p-eEF2 after adding PQBP1 (n = 4) or PQBP1D45–81 (n = 3) in the in vitro eEF2 phosphorylation assay. The data are represented as mean ± SD. (B) Overexpression of PQBP1 suppresses eEF2 phosphorylation in SK-N-BE(2) cells. Bottom: quantification of relative protein levels is represented as mean ± SD of three independent experiments. eEF2, p = 0.51; p-eEF2, p = 0.0047; ns, p > 0.05; **p % 0.01; two-tailed unpaired Student’s t test. See also Figures S3 and S4.

Article Snippet: Briefly, 6 mg eEF2 was incubated with 9 mg eEF2K in 30 mM HEPES-KOH pH 7.5, 150 mM CaCl2, 10 mM MgAC2, 5 mM DTT, 0.5 mM ATP at 30 C for 30 min. eEF2K inhibitor TX1918 (Santa Cruz) was used as a negative control.

Techniques: Phospho-proteomics, In Vitro, Western Blot, Staining, Over Expression, Two Tailed Test