Comparison of two reducing agents dithiothreitol and tris(3-hydroxypropyl)phosphine for in vitro kinetic assay of vitamin K epoxide reductase.
Comparison of two reducing agents dithiothreitol and tris(3-hydroxypropyl)phosphine for in vitro kinetic assay of vitamin K epoxide reductase.
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两种还原剂二硫苏糖醇和三(3-羟丙基)膦用于维生素 K 环氧化物还原酶体外动力学测定的比较。
DOI：
10.1016/j.vas.2020.100095
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发表时间：
2020
期刊：
影响因子：
--
通讯作者：
Mayumi Ishizuka.
中科院分区：
文献类型：
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作者：
Kazuki Takeda;Ayuko Morita;Yoshinori Ikenaka;Shouta M.M.Nakayama;Mayumi Ishizuka.
维生素K环氧化物还原酶（VKOR）是抗凝血剂（如华法林）的靶酶，可用作药物或灭鼠剂。需要评估VKOR活性，以确保这些药物的正确使用。二硫苏糖醇（DTT）是一种典型的二硫化物还原剂，用作体外VKOR测定的底物。然而，DTT被认为是有问题的，因为它的副作用。三（3-羟丙基）膦（THP）已被发现是一个可靠的替代DTT，所示的VKOR与它们的动力学分析。THP的Vmax和Km值均显著低于DTT，但二者对华法林的Vmax/Km和IC 50无显著性差异。
Vitamin K epoxide reductase (VKOR) is a target enzyme for anticoagulants, such as warfarin, that are used as medicines or rodenticides. Assessing VKOR activity is required to ensure the proper usage of these drugs. Dithiothreitol (DTT) is a typical disulfide reductant that is used as a substrate forin vitroVKOR assays. However, DTT is considered problematic because of its side effects. Tris(3-hydroxypropyl)phosphine (THP) has been found to be a reliable alternative to DTT, as shown by kinetic analyses of the VKOR with them. THP showed significantly lowerVmaxandKmvalues than those of DTT; however, there was no significant difference in theirVmax/Kmand IC50for warfarin.