Stabilization of warfarin-binding pocket of VKORC1 and VKORL1 by a peripheral region determines their different sensitivity to warfarin inhibition.
Stabilization of warfarin-binding pocket of VKORC1 and VKORL1 by a peripheral region determines their different sensitivity to warfarin inhibition.
复制标题
外周区域对 VKORC1 和 VKORL1 华法林结合袋的稳定决定了它们对华法林抑制的不同敏感性
DOI：
10.1111/jth.14127
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发表时间：
2018-06
期刊：
影响因子：
--
通讯作者：
Li W
中科院分区：
文献类型：
--
作者：
Shen G;Li S;Cui W;Liu S;Liu Q;Yang Y;Gross M;Li W
人类基因组编码维生素K-环氧化物还原酶的两个旁系同源物，VKORC 1和VKORL 1，支持血液凝固和其他维生素K依赖性过程。华法林抑制这两种酶，但VKORL 1对华法林相对耐药。了解VKORL 1和VKORC 1之间的差异，以及VKORC 1中华法林耐药（WR）突变的原因。我们进行了系统的诱变和分析华法林的反应与基于细胞的活性测定。质谱分析用于检测细胞的氧化还原状态。VKORC 1和VKORL 1采用类似的细胞内氧化还原状态，具有四跨膜螺旋拓扑结构。在VKORC 1中鉴定的大多数WR突变也赋予VKORL 1抗性，表明华法林在共同的结合位点抑制这些旁系同源物。一组WR突变，远离华法林结合位点，在VKORL 1比VKORC 1中显示出显著更低的耐药性，这意味着它们不同的华法林反应是由外周相互作用决定的。值得注意的是，我们确定了一个关键的外围区域，其中单个突变，Glu 37 Lys或His 46 Tyr，大大增加了华法林敏感性的VKORL 1。在这些华法林敏感的VKORL 1突变体的背景下，野生型VKORL 1中表现出相对较低抗性的WR突变变得更具抗性，这表明结构转换类似于VKORC 1。在该外周区域，我们还鉴定了赋予VKORL 1的华法林敏感性的人类SNP。VKORC 1和VKORL 1的外周区域主要维持其共同的华法林结合口袋的稳定性，这种相互作用的差异决定了它们对华法林抑制的相对敏感性。这个新模型也解释了大多数位于VKORC 1外围区域的WR突变。
The human genome encodes two paralogs of vitamin-K-epoxide reductase, VKORC1 and VKORL1, that support blood coagulation and other vitamin-K-dependent processes. Warfarin inhibits both enzymes, but VKORL1 is relatively resistant to warfarin. To understand the difference between VKORL1 and VKORC1, and the cause of warfarin-resistant (WR) mutations in VKORC1. We performed systematic mutagenesis and analyzed warfarin responses with a cell-based activity assay. Mass spectrometry analyses were used to detect cellular redox state. VKORC1 and VKORL1 adopt a similar intracellular redox state with four-transmembrane-helix topology. Most WR mutations identified in VKORC1 also confer resistance in VKORL1, indicating that warfarin inhibits these paralogs at a common binding site. A group of WR mutations, distant from the warfarin-binding site, show significantly less resistance in VKORL1 than in VKORC1, implying that their different warfarin responses are determined by peripheral interactions. Remarkably, we identify a critical peripheral region in which single mutations, Glu37Lys or His46Tyr, drastically increase the warfarin sensitivity of VKORL1. In the background of these warfarin-sensitive VKORL1 mutants, WR mutations showing relative less resistance in wild-type VKORL1 become much more resistant, suggesting a structural conversion to resemble VKORC1. At this peripheral region, we also identified a human SNP that confers warfarin sensitivity of VKORL1. Peripheral regions of VKORC1 and VKORL1 primarily maintain the stability of their common warfarin-binding pocket, and differences of such interactions determine their relative sensitivity to warfarin inhibition. This new model also explains most WR mutations located at the peripheral regions of VKORC1.
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影响因子：
16.8
作者：
Shen G;Cui W;Zhang H;Zhou F;Huang W;Liu Q;Yang Y;Li S;Bowman GR;Sadler JE;Gross ML;Li W
通讯作者：
Li W
影响因子：
64.8
作者：
Rost, S;Fregin, A;Oldenburg, J
通讯作者：
Oldenburg, J
影响因子：
3.5
作者：
Gebauer, Markus
通讯作者：
Gebauer, Markus
影响因子：
16.8
作者：
Czogalla, Katrin J.;Biswas, Arijit;Oldenburg, Johannes
通讯作者：
Oldenburg, Johannes
影响因子：
2.9
作者：
FASCO, MJ;PRINCIPE, LM;FRIEDMAN, PA
通讯作者：
FRIEDMAN, PA