Chemical and immunological properties of reduced and alkylated polypeptide chains of bovine fibrinogen.
Chemical and immunological properties of reduced and alkylated polypeptide chains of bovine fibrinogen.
复制标题
牛纤维蛋白原还原和烷基化多肽链的化学和免疫学特性。
DOI：
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发表时间：
1972
期刊：
影响因子：
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通讯作者：
H. Furthmayr
中科院分区：
文献类型：
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作者：
R. Gollwitzer;R. Timpl;U. Becker;H. Furthmayr
通过cm -纤维素层析从还原和羧甲基化的牛纤维蛋白原中分离出α、β和γ链。电泳纯多肽链可通过三种不同的方法进行判断。这些链在pH为8或更高的缓冲液中可溶，但表现出非共价聚集。在十二烷基硫酸盐-聚丙烯酰胺凝胶电泳中，α链、β链和γ链的分子量分别为62000、58000和48000。每条链在氨基酸组成和CNBr处理后获得的片段模式上都有很大的不同。未解析的α/β物质的出现表明单个链的微观异质性。至少对于α链，亚硫酸盐裂解产生的产物纯度较低。
The α, β and γ chains were isolated from reduced and carboxymethylated bovine fibrinogen by chromatography on CM-cellulose. Electrophoretically pure polypeptide chains could be obtained as judged by three different methods. The chains were soluble in buffers at or above pH 8 but exhibited non-covalent aggregation. The molecular weights of the α, β and γ chains were estimated in dodecylsulfate-polyacrylamide gel electrophoresis as 62000, 58000 and 48000, respectively. Each chain differed considerably from the other in amino acid composition as well as in the fragment pattern obtained after CNBr treatment. The occurrence of unresolved α/β material suggested microheterogeneity of the individual chains. Sulfitolysis yielded, at least for the α chain, less pure products.
Between 20% and 54% of the antibodies in rabbit antisera to native fibrinogen precipitated with the α and/or β chain. Absorption studies, gel precipitation and hemagglutination-inhibition suggested a considerable antigenic homology of both chains. They could be clearly distinguished from the γ chain which reacted poorly in precipitation test but considerably better in passive hemagglutination. Cyanogen bromide cleavage did not destroy the serologic activity of the chains. These findings indicated that a considerable number of the antigenic determinants on fibrinogen are not dependent on intact conformation. However, antibodies specifically reacting with the native molecule occur as well.