Identification of proNeuropeptide FFA peptides processed in neuronal and non-neuronal cells and in nervous tissue.
Peptides which should be generated from the neuropeptide FF (NPFF) precursor were identified in a neuronal (human neuroblastoma SH-SY5Y) cell line and in COS-7 cells after transient transfection of the human proNPFFA cDNA and were compared with those detected in the mouse spinal cord. After reverse-phase high performance liquid chromatography of soluble material, NPFF-related peptides were immunodetected with antisera raised against NPFF and identified by using on-line capillary liquid chromatography/nanospray ion trap tandem mass spectrometry. Neuronal and non-neuronal cells generated different peptides from the same precursor. In addition to NPFF, SQA-NPFF (Ser-Gln-Ala-Phe-Leu-Phe-Gln-Pro-Gln-Arg-Phe-amide) and NPAF were identified in the human neuroblastoma while only NPFF was clearly identified in COS-7 cells. In mouse, in addition to previously detected NPFF and NPSF, SPA-NPFF (Ser-Pro-Ala-Phe-Leu-Phe-Gln-Pro-Gln-Arg-Phe-amide), the homologous peptide of SQA-NPFF, were characterized. These data on intracellular processing of proNeuropeptide FFA are discussed in regard to the known enzymatic processing mechanisms.
D000818:Animals; D019556:COS Cells; D002851:Chromatography, High Pressure Liquid; D006801:Humans; D013058:Mass Spectrometry; D051379:Mice; D009447:Neuroblastoma / Q000378:metabolism; D009842:Oligopeptides / Q000302:isolation & purification*; D010455:Peptides / Q000302:isolation & purification*; D013116:Spinal Cord / Q000378:metabolism
D009842:Oligopeptides; D010455:Peptides; C053651:phenylalanyl-leucyl-phenylalanyl-glutaminyl-prolyl-glutaminyl-arginyl-phenylalaninamide