2 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 o f basic amino acid (aa) residues in the NGF proregion suggests that tw o or three peptides other than NGF may be generated by proteolytic cle avage, Synthetic peptides of 29 aa (LIP1) and 38aa (LIP2) correspondin g to the sequences -71 to -43 and -40 to -3 of the proNGF, respectivel y, were used in this study, ELISA specific for these two peptides reve aled their presence in the rat intestine. LIP1 was localized by immuno histochemistry in endocrine cells of the intestinal epithelium, and LI P2 was immunoprecipitated from an intestinal extract, We also provide evidence for the presence of specific receptors for LIP2 in several ce ll lines, Scatchard analysis indicated the presence of a low affinity binding site with a K-d of similar to 10(-7) M and a high affinity bin ding site of 10(-9) M. Cross-linking studies revealed receptor forms o f 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 proce ss of neurite outgrowth. Furthermore, both propeptides induced rapid t yrosine phosphorylation of the Trk protein in both prostatic adenocarc inoma 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.