Purification of the growth-associated protein GAP-43 by reversed phase chromatography: amino acid sequence analysis and cDNA identification

• Published in: Brain research Vol. 510; no. 2; p. 259
• Main Authors: Changelian, P S, Meiri, K, Soppet, D, Valenza, H, Loewy, A, Willard, M
• Format: Journal Article
• Language: English
• Published: Netherlands 05.03.1990
• Subjects: Amino Acid Sequence; Animals; Base Sequence; Brain - metabolism; Chromatography, High Pressure Liquid; DNA - analysis; GAP-43 Protein; Membrane Glycoproteins - genetics; Membrane Glycoproteins - isolation & purification; Molecular Sequence Data; Nerve Growth Factors - genetics; Nerve Growth Factors - isolation & purification; Nerve Tissue Proteins - genetics; Nerve Tissue Proteins - isolation & purification; Nucleic Acid Hybridization; Rats
• ISSN: 0006-8993
• DOI: 10.1016/0006-8993(90)91376-R

GAP-43 is a neuronal phosphoprotein. Increased synthesis and axonal transport of GAP-43 has been associated with axon growth, and altered phosphorylation of GAP-43 has been associated with changes in synaptic efficacy. Here we report a rapid and effective procedure employing reverse-phase HPLC for the purification of GAP-43 from rat brain. To characterize the protein purified by this procedure, we generated proteolytic fragments and determined their amino acid sequences. These directly determined sequences, corresponding to 56% of the GAP-43 amino acids, confirm recently reported sequences deduced from the nucleotide sequences of cDNAs. Using oligonucleotide probes constructed according to these amino acid sequences, we identified GAP-43 cDNAs in a library prepared from neonatal rat superior cervical ganglion cells. One of these cDNAs was 1.1 kB in size; it hybridized specifically with a 1.5 kB RNA from brain, but not from liver, and contained the entire coding sequence for GAP-43. This cDNA differed from recently reported cDNAs in its 3' untranslated region.