Mandle R J, Colman R W, Kaplan A P
Proc Natl Acad Sci U S A. 1976 Nov;73(11):4179-83. doi: 10.1073/pnas.73.11.4179.
Prekallikrein and high-molecular-weight kininogen were found associated in normal human plasma at a molecular weight of 285,000 as assessed by gel filtration on Sephadex G-200. The molecular weight of prekallikrein in plasma that is deficient in high-molecular-weight kininogen was 115,000. This prekallikrein could be isolated at a molecular weight of 285,000 after plasma deficient in high-molecular-weight kininogen was combined with plasma that is congenitally deficient in prekallikrein. Addition of purified 125I-labeled prekallikrein and high-molecular-weight kininogen to the respective deficient plasma yielded a shift in the molecular weight of prekallikrein, and complex formation could be demonstrated by incubating prekallikrein with high-molecular weight kininogen. This study demonstrates that prekallikrein and high-molecular-weight kininogen are physically associated in plasma as a noncovalently linked complex and may therefore be adsorbed together during surface activation of Hageman factor. The complex is disrupted when these proteins are isolated by ion exchange chromatography.
通过在葡聚糖凝胶G - 200上进行凝胶过滤评估,发现前激肽释放酶和高分子量激肽原在正常人血浆中以分子量285,000的形式存在。在高分子量激肽原缺乏的血浆中,前激肽释放酶的分子量为115,000。在高分子量激肽原缺乏的血浆与先天性前激肽释放酶缺乏的血浆混合后,可以分离出分子量为285,000的这种前激肽释放酶。将纯化的125I标记的前激肽释放酶和高分子量激肽原添加到各自缺乏的血浆中,导致前激肽释放酶分子量发生变化,并且通过将前激肽释放酶与高分子量激肽原一起孵育可以证明复合物的形成。这项研究表明,前激肽释放酶和高分子量激肽原在血浆中以非共价连接的复合物形式物理结合,因此在接触因子表面激活过程中可能会一起被吸附。当通过离子交换色谱法分离这些蛋白质时,复合物会被破坏。