Research on the primitive complement system in agnathan cyclostomes

无颌环口动物原始补体系统的研究

• 批准号: 03640631
• 负责人: FUJII Tamotsu
• 金额: $ 1.47万
• 依托单位: Hiroshima Women's 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: 1991
• 资助国家: 日本
• 起止时间: 1991 至 1993
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-03640631/
• 关键词: Cyclostome; Hagfish; Primitive complement system; Alternative pathway; Third component of complement (C3); Irregular cleavage of pro-C3; Opsonin; アイソタイプ; 代替(第2)経路; 補体第3成分

A protein from hagfish serum that cross-reacted with the third component of hagfish complement (C3-1) was purified to homogeneity and its structural properties were compared with those of two-subunit-chain (115 kDa and 72 kDa) C3-1. This protein (termed C3-2), when purified from the plasma, consisted of three disulfide-linked polypeptide chains (77 kDa, 72 kDa and 30kDa). On immunoelectrophoresis, purified C3-2 had a more anodal mobility compared to the beta mobility of C3-1. However, immunochmical analysis revealed that C3-2 after the initial step in the purification consisted of two disulfide-linked polypeptide chains (105 kDa and 72 kDa). Treatment of C3-2 with methylamine, prior to the spectrophotometric titration of free SH content in this molecule, did not significantly affect the titration end point, suggesting the disruption of a thiol ester bond in C3-2. Amino acid sequence analysis of the amino-termini of the subunits of C3-2 revealed that 77 amino acid residues at the amino-terminus of native alpha-chain were missing from the 77-kDa as well as 105-kDa polypeptides from C3-2. These results indicate that C3-2 isolated in this study is analogous to C3b. Furthermore, sequencing data indicate that most of the native C3 from hagfish occur in the serum with an irregular two-subunit (alpha + gamma and beta) chain structure, as a result of the one-sided processing of putative hagfish pro-C3 at the beta-alpha processing site exclusively, and, moreover that only the molecular features of degenerated hagfish C3 (C3-2) are altered during its purification to generate a three-chain structure

从鳗血清中提纯了一种与鳗鱼补体第三组分(C3-1)发生交叉反应的蛋白质，并将其结构性质与两亚单位链(115 kDa和72 kDa)C3-1的结构性质进行了比较。该蛋白(命名为C3-2)从血浆中纯化后，由77 kDa、72 kDa和30 kDa三条二硫键连接多肽链组成。在免疫电泳法上，与C3-1的β迁移率相比，纯化的C3-2具有更强的阳离子迁移率。免疫化学分析表明，初步纯化后的C3-2由两条二硫键连接的多肽链组成，分别为105 kDa和72 kDa。在分光光度滴定C3-2分子中的游离SH含量之前，用甲胺处理C3-2对滴定终点没有显著影响，这表明C3-2中的硫醇酯键被破坏。对C3-2亚基氨基端的氨基酸序列分析表明，C3-2的77 kDa多肽和105 kDa多肽缺失77个氨基酸残基。这些结果表明，本研究分离到的C3-2类似于C3b。此外，测序数据表明，由于在β-α加工位点对假定的八脚鳗原-C3进行了片面处理，因此来自八脚鳗的大部分天然C3存在于血清中，具有不规则的两个亚单位(α+伽马和β)链结构，而且，在纯化过程中，只有退化的八脚鳗C3(C3-2)的分子特征被改变，以产生三链结构

项目成果

Fujii, T.: "Host-defense mechanisms in cyclostomes (in Japanese)" Rinshou Men-eki. 23. 1625-1634 (1991)

Fujii, T.：“环口动物的宿主防御机制（日语）”Rinshou Men-eki。

Holland,N.D.: "Engrailed expression during development of a lamprey,Lampetra japonica:a possible clue to homologies between agnathan and gnathostome muscles of the mandibular arch." Development,Growth & Differentiation. 35. 153-160 (1993)

Holland，N.D.：“七鳃鳗（Lampetra japonica）发育过程中的刻痕表达：下颌弓无颌肌和颌口肌同源性的可能线索。”

Fujii, T.: "Phylogeny of complement system (in Japanese)" Saibou. 23. 543-547 (1991)

Fujii, T.：“补体系统的系统发育（日语）”Saibou。

Fujii, T., T.Nakamura, A.Sekizawa, and S.Tomonaga: "Isolation and characterization of a protein from hagfish serum that is homologous to the third component of the mammalian complement system" Journal of Immunology. 148. 117-123 (1992)

Fujii, T.、T.Nakamura、A.Sekizawa 和 S.Tomonaga：“从盲鳗血清中分离和表征与哺乳动物补体系统第三种成分同源的蛋白质”《免疫学杂志》。

Holland, N.D., L.Z.Holland, Y.Honma, and T.Fujii: "Engrailed expression during development of a lamprey, Lampetra japonica : a possible clue to homologies between agnathan and gnathostome muscles of the mandibular arch" Development, Growth and Differentia

Holland, N.D.、L.Z.Holland、Y.Honma 和 T.Fujii：“七鳃鳗 (Lampetra japonica) 发育过程中的 Engrailed 表达：下颌弓无颌肌和颌口肌同源性的可能线索”发育、生长和分化