Evidence of normal functional levels of activated protein C inhibitor in combined Factor V/VIII deficiency disease.
Evidence of normal functional levels of activated protein C inhibitor in combined Factor V/VIII deficiency disease.
复制标题
复合因子 V/VIII 缺乏症中活化蛋白 C 抑制剂功能正常水平的证据。
DOI：
10.1172/jci110725
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发表时间：
1982
期刊：
影响因子：
--
通讯作者：
Kisiel,W
中科院分区：
文献类型：
--
作者：
Canfield,WM;Kisiel,W
人活化蛋白C（APC）是一种血浆丝氨酸蛋白酶，具有酰胺分解和抗凝活性。APC的酰胺分解和抗凝活性在正常血浆中被中和的速率与从4个合并因子V/VIII缺乏症的个体获得的血浆中观察到的速率基本相同。放射性碘标记的APC与正常人血浆或组合的因子V/VIII缺陷的血浆一起孵育导致形成酶和血浆蛋白的稳定复合物（Mr = 96，000），如通过十二烷基硫酸钠-聚丙烯酰胺凝胶电泳测定的。用氟磷酸二异丙酯预处理放射性标记的APC可防止酶-蛋白复合物的形成。基于其与放射性标记的APC形成复合物的能力，通过硫酸铵分级分离、肝素-琼脂糖层析和QAE-Sephadex A-50层析从正常人血浆中纯化APC结合蛋白至均一。APC结合蛋白（Mr = 54，000）是一种糖蛋白，其氨基末端序列为Gly-Arg-Thr-Cys-Pro-Lys-Pro-Asp。APC结合蛋白的氨基端序列与牛初乳抑制剂和胰蛋白酶抑制剂具有相当大的同源性，但与血浆丝氨酸蛋白酶抑制剂没有明显的序列同源性。亲和纯化的抗体对APC结合蛋白免疫沉淀复合物的放射性标记的APC和天然APC结合蛋白从正常人血浆。复合物的形成几乎消除了APC结合蛋白的血浆免疫耗竭。定量电免疫分析表明，正常血浆中APC结合蛋白抗原的水平与4名凝血因子V/VIII联合缺乏症患者的血浆基本相同。图片
Human activated protein C (APC) is a plasma serine protease that possesses amidolytic and anticoagulant activity. The rate at which the amidolytic and anticoagulant activity of APC was neutralized in normal plasma was essentially identical to that observed in plasma obtained from four individuals with combined Factor V/VIII deficiency disease. Incubation of radioiodinated APC with either normal human plasma or the combined Factor V/VIII-deficient plasmas resulted in the formation of a stable complex (Mr = 96,000) of the enzyme and a plasma protein as determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Pretreatment of the radiolabeled APC with diisopropyl fluorophosphate prevented the formation of the enzyme-protein complex. On the basis of its ability to form a complex with radiolabeled APC, the APC-binding protein was purified to homogeneity from normal human plasma by ammonium sulfate fractionation, heparin-agarose chromatography, and QAE-Sephadex A-50 chromatography. The APC-binding protein (Mr = 54,000) is a glycoprotein, and possesses an amino-terminal sequence of Gly-Arg-Thr-Cys-Pro-Lys-Pro-Asp. The amino-terminal sequence of the APC-binding protein exhibited considerable homology with bovine colostrum inhibitor and pancreatic trypsin inhibitor, but no apparent sequence homology with the plasma serine protease inhibitors. Affinity-purified antibody against APC-binding protein immunoprecipitated a complex of radiolabeled APC and native APC-binding protein from normal human plasma. Complex formation was virtually eliminated in plasma immunodepleted of the APC-binding protein. Quantitative electroimmunoassay indicated essentially equal levels of APC-binding protein antigen in normal plasma compared with plasma from four patients with combined Factor V/VIII deficiency disease.Images