Structure,function and movement of insect brain peptides,particularly prothoracicotropic hormone.

昆虫脑肽，特别是促前胸腺激素的结构、功能和运动。

• 批准号: 01060004
• 负责人: SUZUKI Akinori
• 金额: $ 144.64万
• 依托单位: The University of Tokyo
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Specially Promoted Research
• 财政年份: 1989
• 资助国家: 日本
• 起止时间: 1989 至 1992
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-01060004/
• 关键词: Silkworm; Peptide hormone; Amino acid Sequence; Nucleotide sequence; Immunohistochemistry; Disulfide bond; Peptide synthesis; Neurosecretion; 前胸腺刺激ホルモン; ボンビキシン; 遺伝子解析; 立体構造; NMR; DELFIA; 羽化ホルモン; 遺伝子発現; アミノ酸配列分析; モノクローナル抗体

1. The primary structure of Bombyx prothoracicotropic hormone(PTTH)was determined by combination of sequencing of the purified PTTH and molecular cloning of cDNA.Bombyx PTTH consists of two identical peptide chains which are held together by disulfide bond(s),and each chain is composed of 109 amino acid residues and a carbohydrate chain bound to an asparagine at position 41.The biologically active,recombinant PTTH has been obtained by using the Escherichia coli system and the disulfide bond location of PTTH were determined using recombinant PTTH.2. The highly effective method for synthesis of bombyxins,an insulin-family peptide,mainly focusing on the selective formation of three disulfide bonds was established,and bombyxin and its various analogs were synthesized.The tertiary structure of bombyxin and its analogs will be determined by NMR analysis.The cDNAs of bombyxin-related peptide were isolated from Samia cynthia ricini and Agrius convolvuri.3. The cDNAs of eclosion hormone(EH)and pheromone biosynthesis activating neuropeptide (PBAN)has been sequenced.PBAN cDNA encodes,besides PBAN,diapause hormone and three putative amidated peptides.4. The primary structure of Bombyx adipokinetic hormone and the N-terminal sequence of diapause hormone were determined.5. The immunochemical studies revealed PTTH-,bombyxin-and PBAN-producing neurosecretory cells.

1.通过对纯化的PTTH进行序列测定和cDNA分子克隆，确定了PTTH的一级结构。PTTH由两条完全相同的肽链组成，每条肽链由109个氨基酸残基和一条糖链组成，糖链通过二硫键连接在一起，糖链与41位的天冬酰胺相连。利用大肠杆菌系统获得了重组PTTH，并利用重组PTTH对PTTH的二硫键进行了定位.建立了高效合成胰岛素家族肽--家蚕毒素的方法，主要针对三个二硫键的选择性形成，合成了家蚕毒素及其各种类似物，并通过核磁共振分析确定了家蚕毒素及其类似物的三级结构。从蓖麻蚕和旋花中分离出了家蚕毒素相关肽的cDNA。3.对羽化激素（EH）和信息素生物合成激活神经肽（PBAN）的cDNA进行了测序，PBAN cDNA除编码PBAN外，还编码滞育激素和三个推测的酰胺化肽.测定了家蚕脂肪运动激素的一级结构和滞育激素的N端序列.免疫组化研究显示PTTH，蚕毒素和PBAN产生的神经分泌细胞。

项目成果

A.Suzuki: "Insect Neurochemistry and Neurophysiology 1989" Humana Press New Jersey, 4 (1990)

A.Suzuki：“昆虫神经化学和神经生理学 1989”Humana Press 新泽西州，4 (1990)

K.Maruyama: "Synthesis of bombyxin-IV,an insulin-like heterodimeric peptide from the silkworm,Bombyxmori." Peptides. 11. 169-171 (1990)

K.Maruyama：“家蚕素-IV 的合成，一种来自蚕 Bombyxmori 的胰岛素样异二聚肽。”

S.Kiriishi: "Comparison of the in vivo and in vitro effects of bombyxin and prothoracicotropic hormone on prothoracic glands of the silkworm,Bombyxmori." Zool.Sci.9. 149-155 (1992)

S.Kiriishi：“家蚕素和促前胸腺激素对家蚕前胸腺的体内和体外影响的比较。”

H.Kuniyoshi: "N-Terminal modified analogs of C-terminal fragments of PBAN with pheromonotropic activity." Insect Biochem.Molec.Biol.22. 399-403 (1992)

H.Kuniyoshi：“具有信息素活性的 PBAN C 端片段的 N 端修饰类似物。”

J.Ishibashi: "Isolation and identification of adipokinetic hormone of the silkworm,Bombyxmori." Biosci.Biotech.Biochem.56. 66-70 (1992)

J.Ishibashi：“家蚕脂肪运动激素的分离和鉴定。”