Human high molecular weight kininogen. Studies of structure-function relationships and of proteolysis of the molecule occurring during contact activation of plasma.
Human high molecular weight kininogen. Studies of structure-function relationships and of proteolysis of the molecule occurring during contact activation of plasma.
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人类高分子量激肽原。
DOI：
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发表时间：
1979
影响因子：
4.8
通讯作者：
J. Griffin
中科院分区：
文献类型：
--
作者：
D. Kerbiriou;J. Griffin
从正常人血浆中以35%的产率纯化人高Mr激肽原。在十二烷基硫酸钠和巯基乙醇存在下，纯化的高Mr激肽原在聚丙烯酰胺凝胶上呈现均一性，并给出具有表观Mr = 110，000的单一蛋白条带。使用沉降平衡技术，观察到的Mr为108，000 +/-2，000。人血浆激肽释放酶切割高Mr激肽原以释放激肽，并产生不含激肽的双链二硫键连接分子，其含有表观Mr = 65，000的重链和表观Mr = 44，000的轻链。轻链富含组氨酸，对带负电荷的物质具有高亲和力。分离的烷基化轻链定量保留了单链母体分子的促凝血活性。125 I-人高Mr激肽原在加入高岭土引发的接触活化过程中在血浆中发生裂解。这种裂解释放激肽并产生双链二硫键连接的分子，取决于血浆中前激肽释放酶和因子XII（Hageman因子）的存在。在存在或不存在高岭土的情况下，将纯化的血浆激肽释放酶添加到正常血浆或前激肽释放酶或因子XII缺陷的血浆中，导致高Mr激肽原的裂解和激肽形成。
Human high Mr kininogen was purified from normal plasma in 35% yield. The purified high Mr kininogen appeared homogeneous on polyacrylamide gels in the presence of sodium dodecyl sulfate and mercaptoethanol and gave a single protein band with an apparent Mr = 110,000. Using sedimentation equilibrium techniques, the observed Mr was 108,000 +/- 2,000. Human plasma kallikrein cleaves high Mr kininogen to liberate kinin and give a kinin-free, two-chain, disulfide-linked molecule containing a heavy chain of apparent Mr = 65,000 and a light chain of apparent Mr = 44,000. The light chain is histidine-rich and exhibits a high affinity for negatively charged materials. The isolated alkylated light chain quantitatively retains the procoagulant activity of the single-chain parent molecule. 125I-Human high Mr kininogen undergoes cleavage in plasma during contact activation initiated by addition of kaolin. This cleavage, which liberates kinin and gives a two-chain, disulfide-linked molecule, is dependent upon the presence of prekallikrein and Factor XII (Hageman factor) in plasma. Addition of purified plasma kallikrein to normal plasma or to plasmas deficient in prekallikrein or Factor XII in the presence or absence of kaolin results in cleavage of high Mr kininogen and kinin formation.