Open AccessIdentification of proNeuropeptide FF A peptides processed in neuronal and non‐neuronal cells and in nervous tissue
Author(s) -
Bonnard Elisabeth,
BurletSchiltz Odile,
Monsarrat Bernard,
Girard JeanPhilippe,
Zajac JeanMarie
Publication year - 2003
Publication title -
european journal of biochemistry
Language(s) - English
Resource type - Journals
eISSN - 1432-1033
pISSN - 0014-2956
DOI - 10.1046/j.1432-1033.2003.03816.x
Subject(s) - peptide , chemistry , intracellular , complementary dna , amide , transfection , neuropeptide , biochemistry , microbiology and biotechnology , biology , receptor , gene
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 proNPFF A 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.