Rat plasma high-molecular-weight kininogen. A simple method for purification and its characterization.
Rat plasma high-molecular-weight kininogen. A simple method for purification and its characterization.
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大鼠血浆高分子量激肽原。
DOI：
10.1016/s0021-9258(18)88944-x
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发表时间：
1985
期刊：
影响因子：
--
通讯作者：
S. Oh‐ishi
中科院分区：
文献类型：
--
作者：
I. Hayashi;H. Kato;S. Iwanaga;S. Oh‐ishi
高分子量激肽原已从大鼠血浆中分离，在三个步骤中，在一个相对较高的产量。在不存在和存在2-巯基乙醇的情况下，纯化的制剂在十二烷基硫酸钠-聚丙烯酰胺凝胶电泳上得到单一条带，并且表观Mr估计为100，000。在与大鼠血浆激肽释放酶孵育时，大鼠高Mr激肽原产生由重链（Mr = 64，000）和轻链（Mr = 46，000）组成的无激肽蛋白，释放缓激肽。将无激肽的蛋白质S-烷基化，并通过锌螯合Sepharose 6 B柱分离其重链和轻链。大鼠高Mr激肽原及其重链和轻链的氨基酸组成与牛高Mr激肽原及其重链和片段1.2-轻链的氨基酸组成非常相似。大鼠高Mr激肽原轻链中的高组氨酸含量表明在牛高Mr激肽原中存在富含组氨酸的蛋白质区域，尽管该区域不被大鼠血浆激肽释放酶裂解。大鼠高Mr激肽原校正到正常值的延长激活部分凝血活酶时间的布朗-挪威Katholiek大鼠血浆已知是缺乏高Mr激肽原和菲茨杰拉德性状血浆。无激肽蛋白具有与完整的高Mr激肽原相同的校正活性。大鼠高Mr激肽原还使大鼠因子XII和前激肽释放酶的表面依赖性活化加速约10倍，这是由高岭土、硫酸直链淀粉和硫苷脂介导的。这些结果表明，大鼠高先生激肽原是非常相似的人和牛高先生激肽原的生化和功能特性。
High-molecular-weight kininogen has been isolated from rat plasma in three steps in a relatively high yield. The purified preparation gave a single band on sodium dodecyl sulfate-polyacrylamide gel electrophoresis in the absence and presence of 2-mercaptoethanol, and the apparent Mr was estimated as 100,000. On incubation with rat plasma kallikrein, rat high Mr kininogen yielded a kinin-free protein consisting of a heavy chain (Mr = 64,000) and a light chain (Mr = 46,000), liberating bradykinin. The kinin-free protein was S-alkylated, and its heavy and light chains were separated by a zinc-chelating Sepharose 6B column. The amino acid compositions of rat high Mr kininogen and its heavy and light chains were very similar to those of bovine high Mr kininogen and its heavy and fragment 1.2-light chains, respectively. A high histidine content in the light chain of rat high Mr kininogen indicated the presence of a histidine-rich region in this protein as in bovine high Mr kininogen, although this region was not cleaved by rat plasma kallikrein. Rat high Mr kininogen corrected to normal values the prolonged activated partial thromboplastin time of Brown-Norway Katholiek rat plasma known to be deficient in high Mr kininogen and of Fitzgerald trait plasma. The kinin-free protein had the same correcting activity as intact high Mr kininogen. Rat high Mr kininogen also accelerated approximately 10-fold the surface-dependent activation of rat factor XII and prekallikrein, which was mediated with kaolin, amylose sulfate, and sulfatide. These results indicate that rat high Mr kininogen is quite similar to human and bovine high Mr kininogens in terms of biochemical and functional properties.