PURIFICATION AND STRUCTURAL-ANALYSIS OF HIPPOCAMPAL CHOLINERGIC NEUROSTIMULATING PEPTIDE
PURIFICATION AND STRUCTURAL-ANALYSIS OF HIPPOCAMPAL CHOLINERGIC NEUROSTIMULATING PEPTIDE
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DOI：
10.1016/0006-8993(92)90465-l
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发表时间：
1992-02-14
期刊：
影响因子：
2.9
通讯作者：
YAMAMOTO, M
中科院分区：
文献类型：
--
作者：
OJIKA, K;KOJIMA, S;YAMAMOTO, M
海马可溶性组分刺激隔内侧核乙酰胆碱（AcCho）合成。 这种刺激活性是从10-12日龄大鼠海马中纯化的。 在纯化过程中，将活性分离成两个级分，并从一个级分中纯化出先前未报道的肽。 该肽的结构为acetyl-Ala-Ala-Asp-Ile-Ser-Gln-Trp-Ala-Gly-Pro-Leu，命名为海马胆碱能神经刺激肽（hippocampal cholinergic neurostimulating peptide，HCNP）。 合成的HCNP和去乙酰化的HCNP（游离HCNP）刺激AcCho合成的内侧隔核培养，以剂量依赖性的方式，但不是文化的纹状体或脊髓前部。 内侧隔核培养物AcCho合成中HCNP和游离HCNP的平均半最大浓度分别为1.0 +/- 0.3 x 10（-10）M和1.0 +/- 0.6 x 10（-11）M。 亲和纯化的多克隆抗体游离HCNP中和粗海马提取物的活性，以及合成的HCNP和游离HCNP。 这些结果表明，HCNP存在于海马提取物中，并参与中枢神经系统特异性胆碱能神经元的发育。
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.