A synthetic peptide substrate for initiation factor-2 kinases

A peptide P(45-56) corresponding to residues 45-56 (sequence: ILLSELSRRRIR) of eIF-2α was synthesised. It was phosphorylated by both of the well characterised eIF-2α kinases viz.; the heme-controlled repressor (HCR) and the double stranded RNA-dependent inhibitor (dsI). Of four other protein kinases tested only protein kinase C (PKC) phosphorylated P(45-56), with complete dependence on phosphatidylserine. Only the residue corresponding to serine-51 in eIF-2α was phosphorylated by HCR, dsI or PKC. The phosphorylation of the peptide by dsI and the phosphorylation of eIF-2α by dsI or HCR showed sigmoidal kinetics with respect to substrate concentration.