tailieunhanh - Báo cáo khoa học: 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 proNPFFAcDNA and were comparedwith those detected in themouse spinal cord. After reverse-phase high performance liquid chromatogra-phy of soluble material, NPFF-related peptides were im-munodetected with antisera raised against NPFF and identified by using on-line capillary liquid chromatography/ nanospray ion trap tandem mass spectrometry. . | Eur. J. Biochem. 270 4187-4199 2003 FEBS 2003 doi Identification of proNeuropeptide FFA peptides processed in neuronal and non-neuronal cells and in nervous tissue Elisabeth Bonnard Odile Burlet-Schiltz Bernard Monsarrat Jean-Philippe Girard and Jean-Marie Zajac Institut de Pharmacologie et de Biologie Structurale Toulouse France 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. Keywords neuropeptide FF electrospray tandem mass spectrometry precursor processing neuroblastoma. Neuropeptide FF NPFF FLFQPQRFamide is a mammalian amidated neuropeptide originally isolated from bovine brain and characterized as a modulator of endogenous opioid functions 1 2 . Two precursors proNPFFA and proNPFFB encoding peptides possessing the PQRF-amide sequence have been cloned in mammals 3 4 . The proNPFFA precursor at basic proteolytic sites should generate two PQRFamide containing peptides 3 and the