Diverse functional properties of Wilson disease ATP7B variants.
Diverse functional properties of Wilson disease ATP7B variants.
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DOI：
10.1053/j.gastro.2011.12.048
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发表时间：
2012-04
期刊：
影响因子：
29.4
通讯作者：
Lutsenko S
中科院分区：
文献类型：
--
作者：
Huster D;Kühne A;Bhattacharjee A;Raines L;Jantsch V;Noe J;Schirrmeister W;Sommerer I;Sabri O;Berr F;Mössner J;Stieger B;Caca K;Lutsenko S
威尔逊病是一种严重的铜代谢紊乱，由ATP7B突变引起，ATP7B编码铜转运腺苷三磷酸酶。该疾病呈现出可变表型，使诊断过程和治疗复杂化。人们对导致这种疾病不同表型的机制知之甚少。我们分析了来自Wilson病患者的28个ATP7B变异，这些变异影响了不同的功能域；利用杆状病毒表达系统在Sf9细胞中表达基因产物。通过测量催化活性和铜（64Cu）向囊泡的转运来分析蛋白质的功能。我们使用共聚焦激光扫描显微镜研究了在哺乳动物细胞中具有可测量的运输活性并被绿色荧光蛋白标记的ATP7B变异的细胞内定位。具有致病性氨基酸取代的ATP7B突变体，即使在相同的功能域内，其特性也会发生很大的变化。一些变体完全丧失了催化和运输活性，而另一些变体丧失了运输活性，但保留了磷中间体的形成或部分丧失了活性。在哺乳动物细胞中，转运能力变异在稳定性和亚细胞定位方面存在差异。与Wilson病相关的ATP7B变异破坏了该蛋白的运输活性，导致其定位错误，并降低了其稳定性。单次检测不足以准确预测ATP7B变异的影响、其产物的功能和Wilson病的发展。这些发现将有助于我们对基因型-表型相关性和疾病发病机制的理解。
Wilson disease is a severe disorder of copper metabolism caused by mutations in ATP7B, which encodes a copper-transporting adenosine triphosphatase. The disease presents with a variable phenotype that complicates the diagnostic process and treatment. Little is known about the mechanisms that contribute to the different phenotypes of the disease. We analyzed 28 variants of ATP7B from patients with Wilson disease that affected different functional domains; the gene products were expressed using the baculovirus expression system in Sf9 cells. Protein function was analyzed by measuring catalytic activity and copper (64Cu) transport into vesicles. We studied intracellular localization of variants of ATP7B that had measurable transport activities and were tagged with green fluorescent protein in mammalian cells using confocal laser scanning microscopy. Properties of ATP7B variants with pathogenic amino-acid substitution varied greatly even if substitutions were in the same functional domain. Some variants had complete loss of catalytic and transport activity, whereas others lost transport activity but retained phosphor-intermediate formation or had partial losses of activity. In mammalian cells, transport-competent variants differed in stability and subcellular localization. Variants in ATP7B associated with Wilson disease disrupt the protein’s transport activity, result in its mislocalization, and reduce its stability. Single assays are insufficient to accurately predict the effects of ATP7B variants the function of its product and development of Wilson disease. These findings will contribute to our understanding of genotype–phenotype correlation and mechanisms of disease pathogenesis.
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影响因子：
4.8
作者：
Tsivkovskii, R;MacArthurs, BC;Lutsenko, S
通讯作者：
Lutsenko, S
影响因子：
4.8
作者：
Tsivkovskii, R;Eisses, JF;Lutsenko, S
通讯作者：
Lutsenko, S
影响因子：
8.6
作者：
Czlonkowska, Anna;Gromadzka, Grazyna;Chabik, Grzegorz
通讯作者：
Chabik, Grzegorz
DOI：
10.1073/pnas.95.18.10854
发表时间：
1998-09-01
影响因子：
11.1
作者：
Payne, AS;Kelly, EJ;Gitlin, JD
通讯作者：
Gitlin, JD
影响因子：
25.7
作者：
Caca, K;Ferenci, P;Berr, F
通讯作者：
Berr, F