Two peptides derived from the nerve growth factor precursor are biologically active.
This report provides evidence that the proregion of the NGF precursor protein contains two novel bioactive peptides. The presence of pairs of basic amino acid (aa) residues in the NGF proregion suggests that two or three peptides other than NGF may be generated by proteolytic cleavage. Synthetic peptides of 29 aa (LIP1) and 38aa (LIP2) corresponding to the sequences -71 to -43 and -40 to -3 of the proNGF, respectively, were used in this study. ELISA specific for these two peptides revealed their presence in the rat intestine. LIP1 was localized by immunohistochemistry in endocrine cells of the intestinal epithelium, and LIP2 was immunoprecipitated from an intestinal extract. We also provide evidence for the presence of specific receptors for LIP2 in several cell lines. Scatchard analysis indicated the presence of a low affinity binding site with a Kd of approximately 10(-7) M and a high affinity binding site of 10(-9) M. Cross-linking studies revealed receptor forms of about 140 kD and 93 kD in a prostatic adenocarcinoma cell line. LIP1 and LIP2 induced rapid F-actin redistribution in PC12 cells within 2 min of incubation, which suggests a role of LIP1 and LIP2 in the process of neurite outgrowth. Furthermore, both propeptides induced rapid tyrosine phosphorylation of the Trk protein in both prostatic adenocarcinoma cells and PC12 cells, thus implicating trk in their mechanism of action. These results support our hypothesis that two peptides within the NGF precursor protein are biologically active.
Dicou
E
E
Institut National de la Santé et de la Recherche Médicale U298, Centre Hospitalier Universitaire, Angers, France.
Pflug
B
B
Magazin
M
M
Lehy
T
T
Djakiew
D
D
Ferrara
P
P
Nerrière
V
V
Harvie
D
D
D000199:Actins / Q000378:metabolism*; D000595:Amino Acid Sequence; D000818:Animals; D001665:Binding Sites; D004797:Enzyme-Linked Immunosorbent Assay; D006801:Humans; D007150:Immunohistochemistry; D007413:Intestinal Mucosa / Q000737:chemistry; D008969:Molecular Sequence Data; D020932:Nerve Growth Factor / Q000032:analysis / Q000737:chemistry / Q000378:metabolism / Q000494:pharmacology*; D009414:Nerve Growth Factors / Q000032:analysis / Q000737:chemistry / Q000378:metabolism / Q000494:pharmacology*; D016716:PC12 Cells; D010446:Peptide Fragments / Q000032:analysis / Q000737:chemistry / Q000378:metabolism / Q000494:pharmacology*; D010766:Phosphorylation; D019000:Phosphotyrosine / Q000378:metabolism; D011498:Protein Precursors / Q000032:analysis / Q000737:chemistry / Q000378:metabolism / Q000494:pharmacology*; D011518:Proto-Oncogene Proteins / Q000378:metabolism*; D051381:Rats; D017208:Rats, Wistar; D020794:Receptor Protein-Tyrosine Kinases / Q000378:metabolism*; D020917:Receptor, trkA; D017475:Receptors, Nerve Growth Factor / Q000378:metabolism*; D014407:Tumor Cells, Cultured
D000199:Actins; C529741:LIP1 proNGF peptide, human; C529742:LIP2 proNGF peptide, human; C104081:Lip2 protein, rat; D009414:Nerve Growth Factors; D010446:Peptide Fragments; D011498:Protein Precursors; D011518:Proto-Oncogene Proteins; D017475:Receptors, Nerve Growth Factor; D019000:Phosphotyrosine; D020932:Nerve Growth Factor; D020794:Receptor Protein-Tyrosine Kinases; D020917:Receptor, trkA