The molecular mechanism of the activation of prophenoloxidase cascade and roles of the cascade on the insect defense

原酚氧化酶级联激活的分子机制及其在昆虫防御中的作用

• 批准号: 09304075
• 负责人: ASHIDA Masaaki
• 金额: $ 20.29万
• 依托单位: Hokkaido University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (A)
• 财政年份: 1997
• 资助国家: 日本
• 起止时间: 1997 至 2000
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-09304075/
• 关键词: innate immunity; pattern recognition; peptidoglycan recognition; beta-1, 3-glucan recognition protein; prophenoloxidase; spatzle; Toll; フェノール酸化酵素前駆体活性化系; 昆虫; 生体防御; セリンプロテアーゼ; 異物認識; Tollレセプター; 抗菌ペプチド; フェノール酸化酵素; 生体防衛; 血液; クチクル; ドロソマイシン

In the innate immunity of insects three main defense reactions operate, namely a) phagocytosis and encapsulation; 2) the activation of prophenoloxidase cascade and blood doagulation; 3) the induction of anti-fungal and anti-bacterial peptides. Intra-cellular signalling pathways for the induction of anti-fungal and anti-bacterial peptide synthesis have been investigated actively for recent years. However, molecules involved in the recognition of bacterial and fungal cell wall components and the mechanism for the transfer of the recognition signal to cells that synthesize anti-microbial peptides. A receptor, Toll is implicated to work to extra cellular recognition signals to cells and the ligand, spatzle has been shown to be produced from prospatzle by a limited proteolysis. To investigate the protease that is directly responsible for the production of spatzle from prospatzle, we prepared Drosophila prospatzle, prepared anti-spatzle/IgG and homogeneous BAEase that is the active form of a zymogen of protease in prophenoloxidase cascade of the silkworm, Bombyx mori. The BAEEase was shown to hydrolyze a peptide bond of prospatzle resulting in the formation spatzle. This result indicate that prophenoloxidase cascade functions as a recognition system for the induction of anti microbial peptide synthesis.

在昆虫的先天免疫中，三种主要防御反应起作用，即a）吞噬和封装； 2）预测酶级联和血液吞噬的激活； 3）抗真菌和抗菌肽的诱导。近年来已经积极研究了抗真菌和抗菌肽合成的细胞内信号传导途径。然而，涉及识别细菌和真菌细胞壁成分的分子以及将识别信号转移到合成抗微生物肽的细胞的机制。受体TOLL与对细胞的额外细胞识别信号有关，并与配体的额外的细胞识别信号有关，Spatzle已通过有限的蛋白水解从prospatzle产生。为了研究直接负责从prospatzle产生的蛋白酶的蛋白酶，我们准备了果蝇prospatzle，制备了抗骨剂/IgG和同质性baease，这是蛋白酶在预知酶cascade cascade of silknoloxidase cascace of silk nork silk虫，bombyx mori中的活性形式。表明baeease可以水解帕特兹的肽键，从而导致形成刺激。该结果表明，预防氧化酶级联反应是诱导抗微生物肽合成的识别系统。

项目成果

Dan Satoh: "Prophenol oxidase-activating enzyme of the silkworm,Bombyx mori (purification,characterization and cDNA cloning)" J.Biol.Chem.274. 7441-7453 (1999)

Dan Satoh：“蚕 Bombyx mori 的丙酚氧化酶激活酶（纯化、表征和 cDNA 克隆）”J.Biol.Chem.274。

芦田正明: "昆虫の液性および細胞性生体防御におけるプロテアーゼカスケード"生体の科学. 51. 194-201 (2000)

Masaaki Ashida：“昆虫体液和细胞防御中的蛋白酶级联”生物科学 51. 194-201 (2000)。

Ochiai, M., Ashida, M.: "A pattern recognition protein for peptidoglycan: cloning the cDNA and the gene of the silkworm, Bombyx mori"J. Biol. Chem.. 274. 11854-11858 (1999)

Ochiai, M., Ashida, M.：“肽聚糖的模式识别蛋白：克隆蚕 Bombyx mori 的 cDNA 和基因”J.

Asano, T., Ashida, M.: "Cuticular prophenoloxidase of the silkworm, Bombyx mori: Purification and demonstration of its transport from hemolymph"J. Biol. Chem.. 276. 11100-11112 (2001)

Asano, T.，Ashida, M.：“家蚕表皮酚氧化酶原：从血淋巴中的纯化和运输演示”J.

Masanori Ochiai: "A pattern-recognition protein for peptidoglycan (cloning the cDNA and gene of the silkworm,Bombyx mori)" J.Biol.Chem.(in press). (1999)

Masanori Ochiai：“肽聚糖的模式识别蛋白（克隆蚕 Bombyx mori 的 cDNA 和基因）”J.Biol.Chem.（出版中）。