ISOLATION AND PARTIAL DETERMINATION OF PRIMARY STRUCTURE OF A NEUROPEPTIDE OF BOMBYX MORI SHOWING SUMMER-MORPH-PRODUCING HORMONE ACTIVITY IN THE ASIAN COMMA BUTTERFLY,POLYGONIA C-AUREUM.

亚洲逗点蝴蝶 Polygonia C-aureum 中表现出夏季形态产生激素活性的家蚕神经肽的初级结构的分离和部分测定。

基本信息

批准号：
06640881
负责人：
ENDO Katsuhiko
金额：
$ 1.34万
依托单位：
YAMAGUCHI UNIVERSITY
依托单位国家：
日本
项目类别：
Grant-in-Aid for General Scientific Research (C)
财政年份：
1994
资助国家：
日本
起止时间：
1994 至 1995
项目状态：
已结题

项目摘要

A neuropeptide (s) showing the same activity as a summer-morph-producing hormone of the Asian comma butterfly, P.c-aureum, was found to exist in brain-suboesophageal ganglion (Br-SG) extract of the silkmoths, Bombyx mori. The peptide was almost the same molecular size with that of bombyxin activating the prothoracic glands of 4th-instar B.mori larvae in vitro.A Sephadex G-50 fraction of Br-SG extracts of B.mori involving molecules of 3-8 kD was applied on CM-, SP-, DEAE- and QAE- Toyoperal columns at pH 5.6 (or pH 6.9), respectively. The SMPH-active peptide was bound to, but bombyxin was not bound to, anion-exchange columns at pH 6.9.By anion-exchange chromatography with a Mono-Q column, the SMPH-activity was recovered in a different fraction from that of bombyxin. A SMPH-active fraction eluted from an Accell QMA cartridge following a Sephadex G-50 column was applied to a reversed-phase HPLC with a linear gradient of 0-50% acetonitoril/50 min (pH 6.9).The SMPH-activity was recovered in two fractions of 40-45% acetonitoril, where only one major peptide band of 2.5-6 kD was stained by SDS-PAGE.The results indicated that a neuropeptide responsible for SMPH-activity of Br-SG extracts of B.mori adults is different from that of bombyxin. The neuropeptide can be purified further by subjecting SMPH-active preparation to anion-exchange chromatography and SDS-PAGE.In addition, some other results was summarized in this reports. They were obtained using the instruments which were supplied by this grant.

在蚕蛾的脑食管下神经节 (Br-SG) 提取物中发现了一种神经肽，其活性与亚洲逗号蝴蝶 (P.c-aureum) 产生夏季形态的激素相同。该肽与在体外激活 4 龄家蚕幼虫前胸腺的家蚕素的分子大小几乎相同。家蚕 Br-SG 提取物的 Sephadex G-50 级分涉及 3-8 kD 的分子分别在 pH 5.6（或 pH 6.9）下应用于 CM-、SP-、DEAE- 和 QAE- Toyoperal 色谱柱。 SMPH 活性肽在 pH 6.9 时与阴离子交换柱结合，但家蚕素未结合。通过使用 Mono-Q 柱的阴离子交换色谱，在与家蚕素不同的级分中恢复了 SMPH 活性。将通过 Sephadex G-50 柱从 Accell QMA 柱上洗脱的 SMPH 活性级分应用于反相 HPLC，线性梯度为 0-50% 乙腈/50 分钟 (pH 6.9)。SMPH 活性得到恢复在 40-45% 乙腈的两份组分中，只有 2.5-6 kD 的一条主要肽带被 SDS-PAGE 染色。结果表明：负责 B.mori 成虫 Br-SG 提取物 SMPH 活性的神经肽与家蚕素不同。通过对SMPH活性制剂进行阴离子交换层析和SDS-PAGE，可以进一步纯化神经肽。此外，本报告还总结了其他一些结果。它们是使用本次赠款提供的工具获得的。