title: Purification and structural analysis of hippocampal cholinergic neurostimulating peptide.
abstract: Hippocampal soluble fraction stimulates acetylcholine (AcCho) synthesis of medial septal nuclei in explant culture system. This stimulating activity was purified from 10-12-day-old rat hippocampus. During purification, the activity was separated into two fractions and a previously unreported peptide was purified from one fraction. The structure of this novel peptide is acetyl-Ala-Ala-Asp-Ile-Ser-Gln-Trp-Ala-Gly-Pro-Leu and we designated it as hippocampal cholinergic neurostimulating peptide (HCNP). Synthesized HCNP and de-acetylated HCNP (free-HCNP) stimulated AcCho synthesis of medial septal nuclei culture, in a dose-dependent manner, but not cultures of corpus striatum or anterior spinal cord. Mean half-maximal concentrations of HCNP and free-HCNP in AcCho synthesis of medial septal nuclei culture were 1.0 +/- 0.3 x 10(-10) M and 1.0 +/- 0.6 x 10(-11) M, respectively. Affinity purified polyclonal antibody to the free-HCNP neutralized the activity of crude hippocampal extract, as well as synthetic HCNP and free-HCNP. These observations suggested that HCNP was present in the hippocampal extract and was involved in development of specific cholinergic neuron in central nervous system.
authors.affiliation: Second Department of Internal Medicine, Medical School, Nagoya City University, Japan.
authors.forename: K
authors.initials: K
authors.lastname: Ojika
authors.forename: S
authors.initials: S
authors.lastname: Kojima
authors.forename: Y
authors.initials: Y
authors.lastname: Ueki
authors.forename: N
authors.initials: N
authors.lastname: Fukushima
authors.forename: K
authors.initials: K
authors.lastname: Hayashi
authors.forename: M
authors.initials: M
authors.lastname: Yamamoto
mesh_terms: D000595:Amino Acid Sequence; D000596:Amino Acids / Q000032:analysis; D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D002268:Carboxypeptidases; D002846:Chromatography, Affinity; D002918:Chymotrypsin; D046508:Culture Techniques; D008969:Molecular Sequence Data; D009419:Nerve Tissue Proteins / Q000737:chemistry / Q000302:isolation & purification*; D009479:Neuropeptides / Q000737:chemistry / Q000302:isolation & purification*; D010447:Peptide Hydrolases; D051381:Rats; D011919:Rats, Inbred Strains
chemical_list: D000596:Amino Acids; D000911:Antibodies, Monoclonal; D009419:Nerve Tissue Proteins; D009479:Neuropeptides; C075274:hippocampal cholinergic neurostimulating peptide; D002268:Carboxypeptidases; D010447:Peptide Hydrolases; C022633:serine carboxypeptidase; C038220:acylaminoacyl-peptidase; D002918:Chymotrypsin