Phosphorylation site of eukaryotic initiation factor 4E.
Eukaryotic protein synthesis initiation factor 4E (eIF-4E) was labeled in situ with [32P]orthophosphate in cultured HeLa cells and rabbit reticulocytes and purified by affinity chromatography. Tryptic digestion yielded one labeled peptide which contained predominantly serine and lysine. After treatment of the protein with citraconic anhydride to block epsilon-amino groups of lysyl residues, tryptic digestion yielded a labeled peptide whose composition was consistent with the structure Trp-Ala-Leu-Trp-Phe-Phe-Lys-Asn-Asp-Lys-Ser(P)-Lys-Thr-Trp-Gln-Ala-Asn-L eu-Arg, one of the arginyl peptides predicted from the human eIF-4E cDNA sequence. The only serine in this peptide is located at position 53 of eIF-4E. Thus, it is concluded that eIF-4E contains a single site of phosphorylation for an endogenous protein kinase, which is Ser-53 in the human eIF-4E sequence.
D000595:Amino Acid Sequence; D000818:Animals; D001665:Binding Sites; D002846:Chromatography, Affinity; D002949:Citraconic Anhydrides / Q000494:pharmacology; D039561:Eukaryotic Initiation Factor-4E; D006367:HeLa Cells / Q000378:metabolism; D006801:Humans; D008875:Middle Aged; D010446:Peptide Fragments / Q000378:metabolism; D010448:Peptide Initiation Factors / Q000378:metabolism*; D010710:Phosphates / Q000378:metabolism*; D010766:Phosphorylation; D011817:Rabbits; D012156:Reticulocytes / Q000378:metabolism; D012694:Serine / Q000378:metabolism; D014357:Trypsin / Q000378:metabolism
D002949:Citraconic Anhydrides; D039561:Eukaryotic Initiation Factor-4E; D010446:Peptide Fragments; D010448:Peptide Initiation Factors; D010710:Phosphates; D012694:Serine; D014357:Trypsin