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Functional analysis of the second and third complement component (C3) in carp using anti-peptide antibodies.

使用抗肽抗体对鲤鱼第二和第三补体成分 (C3) 进行功能分析。

基本信息

批准号：
10660184
负责人：
NAKAO Miki
金额：
$ 2.3万
依托单位：
KYUSHU UNIVERSITY
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
1998
资助国家：
日本
起止时间：
1998 至 1999
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-10660184/
关键词：
FISH IMMUNOLOGY COMPLEMENT C3 DIVERSITY CLONING PROTEIN 免疫魚類コイ B因子

项目摘要

We describe two diverged B/C2-like cDNA clones (B/C2-A and B/C2-B) isolated from a bony fish, common carp (Cyprinus carpio). B/C2-A shares the same domain structure with B and C2 of the vertebrate reported so far, and shows a close similarity to zebrafish B and medaka fish B/C2. These teleost sequences show almost the same similarity to C2 and B of higher vertebrates. In contrast, B/C2-B has a novel structural feature in that it contains four SCR modules and does not have a close relative upon phylogenetic analysis. Northern blotting revealed the presence of two transcripts with different size for each of B/C2-A and B/C2-B in the hepatopancreas of carp. A possible phylogenetic relationship between two carp sequences, and mammalian C2 and B is discussed. We have also isolated eight distinct cDNA clones encoding C3 from a single carp (Cyprinus carpio). The eight sequences were grouped into five C3 types, designated C3-H1, C-H2, C3-S, C3-Q1, and C3-Q2. On the other hand, five distinct C3 forms, named C3-1 to C3-5, were purified from serum of a single carp. N-terminal sequencing and covalent binding to [3H] glycine identified C3-1as the translated product of C3-S, while C3-2 was that of C3-H1, and C3-5 that of C3-H2. C3-1 showed a hemolytic activity three-fold higher than that of C3-2, whereas C3-5 was inactive, suggesting that the thioester-catalytic mechanism is not a necessary determinant for the C3 activity and that C3 lacking the catalytic histidine plays a significant role in the complement system of carp and probably other bone fish.

我们描述了从硬骨鱼鲤鱼 (Cyprinus carpio) 中分离出的两个不同的 B/C2 样 cDNA 克隆（B/C2-A 和 B/C2-B）。 B/C2-A与迄今为止报道的脊椎动物的B和C2具有相同的结构域结构，并且与斑马鱼B和青鳉鱼B/C2非常相似。这些硬骨鱼序列与高等脊椎动物的 C2 和 B 几乎具有相同的相似性。相比之下，B/C2-B具有一个新颖的结构特征，即它包含四个SCR模块，并且在系统发育分析中没有近亲。 Northern 印迹显示鲤鱼肝胰腺中存在 B/C2-A 和 B/C2-B 两个不同大小的转录本。讨论了两个鲤鱼序列与哺乳动物 C2 和 B 之间可能的系统发育关系。我们还从单一鲤鱼 (Cyprinus carpio) 中分离出八个编码 C3 的不同 cDNA 克隆。这八个序列分为五种 C3 类型，指定为 C3-H1、C-H2、C3-S、C3-Q1 和 C3-Q2。另一方面，从一条鲤鱼的血清中纯化出五种不同的 C3 形式，命名为 C3-1 至 C3-5。 N-末端测序和与[3H]甘氨酸的共价结合鉴定出C3-1是C3-S的翻译产物，而C3-2是C3-H1的翻译产物，C3-5是C3-H2的翻译产物。 C3-1的溶血活性比C3-2高三倍，而C3-5则无活性，这表明硫酯催化机制并不是C3活性的必要决定因素，并且缺乏催化组氨酸的C3在鲤鱼和可能其他骨鱼的补体系统中发挥着重要作用。