Gene analysis of inherited antithrombin deficiency and functional analysis of abnormal antithrombin protein (N87D)
Gene analysis of inherited antithrombin deficiency and functional analysis of abnormal antithrombin protein (N87D)
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遗传性抗凝血酶缺陷基因分析及异常抗凝血酶蛋白（N87D）功能分析
DOI：
10.1007/s12185-017-2352-8
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发表时间：
2018
期刊：
影响因子：
2.1
通讯作者：
Morishita E
中科院分区：
文献类型：
--
作者：
Kamijima S;Sekiya A;Takata M;Nakano H;Murakami M;Nakazato T;Asakura H;Morishita E
遗传性抗凝血酶（AT）缺乏症是临床上最显着的先天性血栓形成倾向之一，遵循常染色体显性遗传模式。我们分析了一名在怀孕期间出现深静脉血栓形成和低 AT 活性的患者的 SERPINC1，并发现了一种新的错义突变 c.259A>G (p.Asn87Asp；N87D)。令人惊讶的是，对父母DNA的分析表明他们不具有这种突变体，因此，这可能是由于新生突变所致。我们还在 COS-1 细胞中表达了这种突变 AT 蛋白，并将其细胞内定位以及细胞内和细胞外抗原水平与野生型 AT 进行了比较。表达实验未发现细胞裂解液中突变型和野生型AT的抗原水平存在显着差异，但COS-1细胞上清液中突变型AT抗原水平明显低于其野生型对应物。免疫荧光没有表明突变型和野生型 AT 在荧光信号的细胞质定位方面有任何差异。我们的研究结果表明，患者的 AT 缺陷可能是由于突变 AT 蛋白 p.Asn87Asp 的细胞外分泌受损所致。
Inherited antithrombin (AT) deficiency is one of the most clinically significant forms of congenital thrombophilia and follows an autosomal dominant mode of inheritance. We analyzed SERPINC1 in a patient who developed deep-vein thrombosis and low AT activity during pregnancy, and identified a novel missense mutation c.259A>G (p.Asn87Asp; N87D). Surprisingly, analysis of the parents’ DNA showed that they did not possess this mutant, and thus, it may have been due to a de novo mutation. We also expressed this mutant AT protein in COS-1 cells and compared its intracellular localization and intracellular and extracellular antigen levels with that of wild-type AT. The expression experiment did not reveal a significant difference in the antigen levels of the mutant and wild-type AT in the cell lysate, but the mutant AT antigen level was markedly lower than that of its wild-type counterpart in the COS-1 cell supernatant. Immunofluorescence did not indicate any difference between the mutant and wild-type AT in terms of cytoplasmic localization of fluorescence signals. Our findings suggest that the patient’s AT deficiency may have been caused by impaired extracellular secretion of mutant AT protein p.Asn87Asp.