Novel pathogenic XK mutations in McLeod syndrome and interaction between XK protein and chorein
Novel pathogenic XK mutations in McLeod syndrome and interaction between XK protein and chorein
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DOI：
10.1212/nxg.0000000000000328
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发表时间：
2019-06-01
影响因子：
3.1
通讯作者：
Sano, Akira
中科院分区：
文献类型：
--
作者：
Urata, Yuka;Nakamura, Masayuki;Sano, Akira
目的检测6例疑似McLeod综合征（MLS）患者的XK基因突变，并探讨ChAc和MLS相关蛋白Chorein与XK蛋白的相互作用。方法采用XK免疫印迹法和舞蹈素免疫印迹法对疑似MLS患者、ChAc携带者、ChAc突变携带者和正常对照者的红细胞膜蛋白进行分析。我们进行了突变分析和XK免疫印迹，以分子诊断疑似MLS的患者。将培养细胞的裂解物与抗XK和抗chorein抗体共免疫沉淀。结果所有疑似MLS病例均被分子诊断为MLS，并发现了新的突变。平均发病年龄为46.8 ± 8岁，比ChAC患者的发病年龄大。免疫印迹分析显示所有MLS患者的舞蹈素免疫反应性显着降低。免疫沉淀分析表明，直接或间接的chorein XK相互作用。结论6例MLS患者均存在XK致病性突变，其中包括新突变。在MLS红细胞膜中，软骨素免疫反应显著降低。此外，我们通过分子分析证明了chorein和XK蛋白之间可能的相互作用。chorein表达的减少与Kell抗原和XK蛋白之间的减少相似，尽管chorein-XK相互作用可能是非共价结合，不像共价Kell-XK复合物。我们的研究结果表明，减少chorein水平缺乏XK蛋白可能与MLS的分子发病机制。
Objective To identify XK pathologic mutations in 6 patients with suspected McLeod syndrome (MLS) and a possible interaction between the chorea-acanthocytosis (ChAc)- and MLS-responsible proteins: chorein and XK protein. Methods Erythrocyte membrane proteins from patients with suspected MLS and patients with ChAc, ChAc mutant carriers, and normal controls were analyzed by XK and chorein immunoblotting. We performed mutation analysis and XK immunoblotting to molecularly diagnose the patients with suspected MLS. Lysates of cultured cells were co-immunoprecipitated with anti-XK and anti-chorein antibodies. Results All suspected MLS cases were molecularly diagnosed with MLS, and novel mutations were identified. The average onset age was 46.8 +/- 8 years, which was older than that of the patients with ChAc. The immunoblot analysis revealed remarkably reduced chorein immunoreactivity in all patients with MLS. The immunoprecipitation analysis indicated a direct or indirect chorein-XK interaction. Conclusions In this study, XK pathogenic mutations were identified in all 6 MLS cases, including novel mutations. Chorein immunoreactions were significantly reduced in MLS erythrocyte membranes. In addition, we demonstrated a possible interaction between the chorein and XK protein via molecular analysis. The reduction in chorein expression is similar to that between Kell antigens and XK protein, although the chorein-XK interaction is a possibly noncovalent binding unlike the covalent Kell-XK complex. Our results suggest that reduced chorein levels following lack of XK protein are possibly associated with molecular pathogenesis in MLS.