Interaction of antibodies to synthetic peptides of proNGF with in vitro synthesized NGF precursors.
Sera raised against three synthetic peptides that reproduce sequences of the pro-nerve growth factor (proNGF) protein were tested in immunoprecipitation experiments using in vitro translation products of SP6-directed NGF mRNA in a rabbit reticulocyte lysate. The interaction of these antibodies with bacterially synthesized chimeric preproNGF was also examined. Digestion of the translation products by the gamma-subunit generated the 22 and 18 kDa intermediates. A predominant 13 kDa intermediate was obtained after digestion of translation products in wheat germ extract. This is shown to be the N-terminal peptide by immunoprecipitation with an anti-peptide serum. These antibodies may be used to detect NGF precursor cleavage products in vivo.
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INSERM U 298, CHRU, Angers, France.
D000818:Animals; D000936:Antigen-Antibody Complex; D003001:Cloning, Molecular; D004247:DNA / Q000235:genetics; D051379:Mice; D020932:Nerve Growth Factor; D009414:Nerve Growth Factors / Q000138:chemical synthesis / Q000235:genetics* / Q000276:immunology; D014176:Protein Biosynthesis; D011498:Protein Precursors / Q000138:chemical synthesis / Q000235:genetics* / Q000276:immunology; D012333:RNA, Messenger / Q000235:genetics; D011817:Rabbits; D012156:Reticulocytes / Q000378:metabolism
D000936:Antigen-Antibody Complex; D009414:Nerve Growth Factors; D011498:Protein Precursors; D012333:RNA, Messenger; C489226:pro-nerve growth factor, mouse; D004247:DNA; D020932:Nerve Growth Factor