Mechanism of Hemolymph Coagulation System in Invertebrates

无脊椎动物血淋巴凝固系统的机制

基本信息

批准号：
02454539
负责人：
IWANAGA Sadaaki
金额：
$ 4.48万
依托单位：
Kyushu University
依托单位国家：
日本
项目类别：
Grant-in-Aid for General Scientific Research (B)
财政年份：
1990
资助国家：
日本
起止时间：
1990 至 1991
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-02454539/
关键词：
Horseshoe crab Blood coagulation Factor B Factor G Serine protease beta-1, 3-glucan Bacterial endotoxin cDNA cloning 体液凝固因子無脊椎動物真菌類カブトガニ血球 Proclotting enzyme IX因子トロンビンタキプレシンゲノム解析抗菌性ペプチド

项目摘要

Limulus proclotting enzyme and factor b associated with endotoxin-sensitive coagulation cascade : novel serine protease zymogens with a new type of "disulfide-knotted domain"Horseshoe crab (Limulus) hemolymph responds to bacterial endotoxin and results in rapid coagulation. This reaction is composed of a cascade consisting of three serine protease zymogens (factor C, factor B, and proclotting enzyme) and a clottable protein (coagulogen), all of which are released by degranulation of the hemocytes on the stimutation of endotoxin. In the present paper, we describe the structures of proclotting enzyme and factor B. CDNA clonings of all the factors associated with this cascade were completed by this study. Proclotting enzyme is a single-chain glycoprotein with molecular mass of 54 kDa. Upon activation by factor B, it is converted to a two-chain active form clotting enzyme composed of an L (25 kDa) and a H (31 kDa) chains. A CDNA for proclotting enzyme encodes a sequence comprising 29 amino … More acid residues of prepro-sequence and 346 residues of the mature protein. Factor B (64 kDa) is also a glycoprotein that exists as a single-chain form and a two-chain zymogen form with an L (25 kDa) and a H (40 kDa) chains. Factor B zymogen is activated to B by active factor C, which is derived from the zymogen factor C that is sensitive to endotoxin. Factor B has 375 amino acid residues in the mature form, in addition to 25 residues of signal sequence. The entire amino acid sequences of the two proteins are similar, suggeting that these proteins were arisen from a gene duplication. Particularly, the sequence identity of serine protease domains at their C-terminus is calculated to be 43.9%. The N-terminal regions up to 60th residue of both proteins share a similar structure with six cysteines that has not been found in any other proteins. The disulfide locations determined on proclotting enzyme indicate that this region consists of a compact "disulfide-knotted domain". The sequence accompanied with the activation of proclotting enzyme was proven to be an Arg-Ile bond, whereas that of factor B, deduced from sequence alignments with other serine proteases, contained a unique-Arg-Gly-Ile-sequence. Less

与内毒素敏感的凝血级联反应的limulus propting酶和因子B：新型的丝氨酸蛋白Zymogens具有新型的“二硫化二硫键结域”类型的马蹄蟹（Limulus）血糖对细菌内毒素的反应，并导致快速凝结。该反应由级联反应组成，该级联反应由三种丝氨酸蛋白酶原（因子C，因子B和proclotting酶）和可蛋白质蛋白（凝结酶）组成，所有这些蛋白质（凝结剂）均通过对内毒素刺激的半细胞脱粒而释放。在本文中，我们描述了与该级联相关的所有因素的prot酶和因子B的结构。 proclotting酶是一种单链糖蛋白，分子质量为54 kDa。通过因子B激活后，将其转换为由L（25 kDa）和A H（31 kDa）链组成的两链活动形式凝结酶。用于protting酶的cDNA编码包含29个氨基的序列…更多的酸保留前序列和346个成熟蛋白的保留。因子B（64 kDa）也是一种糖蛋白，它作为单链形式和具有L（25 kDa）和A H（40 kDa）链的两链Zymogen形式存在。通过活性因子C激活B酶原因子C，该因子C源自对内毒素敏感的酶原因子C。因子B除了25个信号序列的残留物外，还具有375个氨基酸以成熟形式保留。两种蛋白质的整个氨基酸序列都是相似的，这表明这些蛋白质是由基因复制引起的。特别是，串行蛋白结构域在其C末端的序列身份计算为43.9％。两种蛋白质的N末端区域的N末端区域具有相似的结构，与六个半胱氨酸相似，在任何其他蛋白质中尚未发现。确定protting酶的二硫化物位置，表明该区域由紧凑的“二硫键打结结构域”组成。伴有proclotting酶激活的序列被证明是Arg-Eile键，而从与其他序列蛋白的序列对齐得出的因子B的序列含有一个独特的Arg-Gly-Gly-ei-Ele-sequencence。较少的