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Isolation of the first component (C1) and the second component (C2) of carp complement

鲤鱼补体第一成分(C1)和第二成分(C2)的分离

基本信息

批准号：
61560214
负责人：
YANO Tomoki
金额：
$ 1.09万
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 (C)
财政年份：
1986
资助国家：
日本
起止时间：
1986 至 1987
项目状态：
已结题

项目摘要

The first component of complement(C1) was isolated from carp serum by a 6-step purification procedure. About 2.8 mg of carp Cl was obtained from 300 ml of carp serum. The final preparatin contained 18% of the initial Cl hemolytic activity and 0.023% of the initial serum prote in, representing 788-fold purification. The high purity of the isolated carp Cl was established by immunoelectrophoresis against anti-whole carp serum (rabbit). The molecular weight of carp C1 was estimated to be 1,020,000. The hemolytic activity of carp C1 was inhibited by EDTA treatment, but this inhibition was overcome by dialysis against a Ca^<2+>-containing buffer. The hemolytic activity of carp C1 was destroyed by heating (50 C, 15 min) or incubation with carrageenan, but was retained when incubated with ammonia, hydrazine or zymosan. These results indicate that carp C1 resembles mammalian C1 in its chemical properties. An intermediate complex EAC1,4,2 was prepared by incubating EAC1,4 (Yano et al., 1986) with hydrazine treated carp serum (Hd), which is deficient in C3, in the presence of Mg^<2+>. The difference between EAC1,4,2 and EAC1,4 was shown by the tests with various inactivated sera. It was also confirmed that the reaction between EAC1,4,2 and C3 proceeds without any divalent cation. The author is now making efforts to isolate C2 from carp serum. Data will be published in the near future.

用6步纯化的方法从鲤鱼血清中分离出补体第一组分(C_1)。从300ml鲤鱼血清中可得到约2.8 mg的鲤鱼氯。最终的制备方法含有初始氯溶血活性的18%和初始血清蛋白的0.023%，纯化倍数为788倍。用抗全鱼血清(兔)的免疫电泳法获得高纯度的鲤鱼氯蛋白。鲤鱼C1的分子量估计为1,020,000。鲤鱼C1的溶血活性可被EDTA处理所抑制，但这种抑制可被含钙缓冲液的透析所克服。加热(50℃，15min)或与卡拉胶孵育可破坏鲤鱼C1的溶血活性，但与氨水、联氨或酵母多糖孵育可保持其溶血活性。这些结果表明鲤鱼c1的化学性质与哺乳动物c1相似。将EAC1，4(Yano等人，1986)与缺乏补体C3的肼处理鲤鱼血清(HD)在Mg~(2+)、lt；2~+和Gt~(2+)存在下孵育，得到了中间体EAC1，4，2。用不同灭活血清进行的试验表明，EAC1、4、2与EAC1、4之间存在差异。还证实了EAC1，4，2和C3之间的反应是在没有任何二价阳离子的情况下进行的。作者目前正在努力从鲤鱼血清中分离C2。数据将在不久的将来公布。