Protein C (PC) plays an important role in the balance of coagulation and anticoagulation. Thus, the detection of PC activity is diagnostically significant for patients with cardiovascular diseases. Presently, the key methods to detect PC activity are the chromogenic assay and activated partial thromboplastin time (APTT) test. PROTAC used in the chromogenic assay is isolated from Agkistrodon contortrix venom as protein C activator (PCA). However, the use of the chromogenic assay is limited because of the high price of PROTAC. In this study, PCA was successfully purified from Agkistrodon acutus venom (AAV) by ion-exchange and gel chromatography. PCA from AAV has a relative molecular mass of 24 kD, calculated from the measurement of 10% sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). The components of PCA were identified by MALDI-TOF/TOF-MS and mascot searches revealed that the coverage rate between PCA and zinc metalloproteinase AaPA from AAV was 21%. The chromogenic assay and APTT test were used to measure the enzymatic activity of PCA, and the results showed that PCA from AAV could specifically activate PC. In summary, the chromogenic assay described herein is highly sensitive and easy to perform.

蛋白C（PC）在凝结和抗凝的平衡中起着重要作用。因此，对于患有心血管疾病的患者，检测PC活性具有重要的诊断意义。目前，检测PC活性的关键方法是生色测定和活化的部分凝血活酶时间（APTT）测试。发色测定中使用的PROTAC是从Agkistrodon contortrix蛇毒中分离出的C蛋白激活剂（PCA）。然而，由于PROTAC的高价，发色测定法的使用受到限制。在这项研究中，成功​​地从Agkistrodon acutus中纯化了PCA。离子交换和凝胶色谱分析毒液（AAV）。AAV的PCA的相对分子量为24 kD，由10％十二烷基硫酸钠-聚丙烯酰胺凝胶电泳（SDS-PAGE）的测量得出。通过MALDI-TOF / TOF-MS鉴定PCA的成分，吉祥物搜索显示PCA与AAV的锌金属蛋白酶AaPA的覆盖率达到21％。用显色法和APTT法测定了PCA的酶活性，结果表明AAV的PCA可以特异性激活PC。总之，本文描述的生色测定法是高度灵敏的并且易于执行。