Human cathepsins K, L, and S: Related proteases, but unique fibrinolytic activity.
Human cathepsins K, L, and S: Related proteases, but unique fibrinolytic activity.
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DOI：
10.1016/j.bbagen.2018.06.015
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发表时间：
2018-09
期刊：
影响因子：
--
通讯作者：
Platt MO
中科院分区：
文献类型：
--
作者：
Douglas SA;Lamothe SE;Singleton TS;Averett RD;Platt MO
纤维蛋白的形成和溶解归因于包括凝血酶激活和纤溶酶蛋白分解在内的蛋白酶激活和纤维蛋白分解的级联反应。半胱氨酸组织蛋白是心血管疾病和糖尿病期间由血管内皮细胞和其他细胞分泌的一种强大的蛋白酶。它们的纤溶活性和止血作用尚未得到很好的描述。将纤维蛋白凝胶聚合，并与重组人组织蛋白(CAT)K、L或S或纤溶酶孵育，进行剂量和时间依赖的研究。对纤维蛋白凝胶的溶解进行成像。用十二烷基硫酸钠-PAGE分析从纤维蛋白凝胶中释放的裂解片段和残存的不溶性纤维蛋白凝胶，在电泳法之前溶解以评估纤维蛋白α，β和γ多肽被组织蛋白酶的降解。复合组织蛋白酶酶谱测定了剩余的有效组织蛋白酶数量。与组织蛋白酶孵育后，α和β纤维蛋白多肽显著丢失，24小时后，猫纤维蛋白凝胶完全溶解。与纤维蛋白的结合稳定了CATL的激活时间；它与多种大小的被切割的纤维蛋白片段有关。在CatK或S中没有观察到这一点。猫在纤维蛋白孵育过程中也保持较长时间的活跃，但其结合/结合不经得起SDS-PAGE的准备。人组织蛋白酶K、L和S具有纤溶活性，可以特异性地降解α和β纤维蛋白多肽链，产生纤溶酶所特有的片段。组织蛋白酶K、L和S纤溶活性的证明导致进一步研究在破坏血管止血或必须考虑非纤溶酶介导的纤溶作用的基于纤维蛋白的工程化血管结构中的贡献作用。
Fibrin formation and dissolution are attributed to cascades of protease activation concluding with thrombin activation, and plasmin proteolysis for fibrin breakdown. Cysteine cathepsins are powerful proteases secreted by endothelial cells and others during cardiovascular disease and diabetes. Their fibrinolytic activity and putative role in hemostasis has not been well described. Fibrin gels were polymerized and incubated with recombinant human cathepsins (cat) K, L, or S, or plasmin, for dose-dependent and time-dependent studies. Dissolution of fibrin gels was imaged. SDS-PAGE was used to resolve cleaved fragments released from fibrin gels and remnant insoluble fibrin gel that was solubilized prior to electrophoresis to assess fibrin α, β, and γ polypeptide hydrolysis by cathepsins. Multiplex cathepsin zymography determined active amounts of cathepsins remaining. There was significant loss of α and β fibrin polypeptides after incubation with cathepsins, with catS completely dissolving fibrin gel by 24 hours. Binding to fibrin stabilized catL active time; it associated with cleaved fibrin fragments of multiple sizes. This was not observed for catK or S. CatS also remained active for longer times during fibrin incubation, but its association/binding did not withstand SDS-PAGE preparation. Human cathepsins K, L, and S are fibrinolytic, and specifically can degrade the α and β fibrin polypeptide chains, generating fragments unique from plasmin. Demonstration of cathepsins K, L, and S fibrinolytic activity leads to further investigation of contributory roles in disrupting vascular hemostasis, or breakdown of fibrin-based engineered vascular constructs where non-plasmin mediated fibrinolysis must be considered.
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影响因子：
2.9
作者：
Keegan PM;Surapaneni S;Platt MO
通讯作者：
Platt MO
DOI：
10.1161/hypertensionaha.113.02141
发表时间：
2014-03
期刊：
Hypertension (Dallas, Tex. : 1979)
影响因子：
--
作者：
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通讯作者：
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影响因子：
37.8
作者：
Lutgens, E;Lutgens, SPM;Cleutjens, KBJM
通讯作者：
Cleutjens, KBJM
影响因子：
2.8
作者：
Liu, Yingxian;Li, Xiangping;Peng, Daoquan;Tan, Zheng;Liu, Hongmin;Qing, Yingnan;Xue, Yanqiong;Shi, Guo-Ping
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影响因子：
8
作者：
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通讯作者：
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