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Analysis of CLCNKB mutations at dimer‐interface, calcium‐binding site, and pore reveals a variety of functional alterations in ClC‐Kb channel leading to Bartter syndrome
Human Mutation ( IF 3.9 ) Pub Date : 2019-12-24 , DOI: 10.1002/humu.23962
Yohan Bignon 1, 2 , Imene Sakhi 1, 2 , Sara Bitam 1, 2 , Naziha Bakouh 1, 2 , Mathilde Keck 1, 2 , Nadia Frachon 2 , Marc Paulais 1, 2 , Gabrielle Planelles 1, 2 , Jacques Teulon 1, 2 , Olga Andrini 3
Affiliation  

Pathological missense mutations in CLCNKB gene give a wide spectrum of clinical phenotypes in Bartter syndrome type III patients. Molecular analysis of the mutated ClC‐Kb channels can be helpful to classify the mutations according to their functional alteration. We investigated the functional consequences of nine mutations in the CLCNKB gene causing Bartter syndrome. We first established that all tested mutations lead to decreased ClC‐Kb currents. Combining electrophysiological and biochemical methods in Xenopus laevis oocytes and in MDCKII cells, we identified three classes of mutations. One class is characterized by altered channel trafficking. p.A210V, p.P216L, p.G424R, and p.G437R are totally or partially retained in the endoplasmic reticulum. p.S218N is characterized by reduced channel insertion at the plasma membrane and altered pH‐sensitivity; thus, it falls in the second class of mutations. Finally, we found a novel class of functionally inactivated mutants normally present at the plasma membrane. Indeed, we found that p.A204T alters the pH‐sensitivity, p.A254V abolishes the calcium‐sensitivity. p.G219C and p.G465R are probably partially inactive at the plasma membrane. In conclusion, most pathogenic mutants accumulate partly or totally in intracellular compartments, but some mutants are normally present at the membrane surface and simultaneously show a large range of altered channel gating properties.

中文翻译:

对二聚体界面,钙结合位点和孔的CLCNKB突变的分析揭示了导致Bartter综合征的ClC-Kb通道的多种功能改变

CLCNKB基因的病理错义突变为III型Bartter综合征患者提供了广泛的临床表型。对突变的ClC-Kb通道进行分子分析可有助于根据突变的功能变化对突变进行分类。我们调查了导致Bartter综合征的CLCNKB基因中的9个突变的功能后果。我们首先确定所有测试的突变都会导致ClC-Kb电流降低。非洲爪蟾的电生理和生化方法相结合卵母细胞和MDCKII细胞中,我们鉴定出三类突变。一类的特征是渠道贩运发生了变化。p.A210V,p.P216L,p.G424R和p.G437R完全或部分保留在内质网中。p.S218N的特征是减少了质膜上的通道插入并改变了pH敏感性;因此,它属于第二类突变。最后,我们发现了通常存在于质膜上的一类新型的功能失活突变体。实际上,我们发现p.A204T改变了pH敏感性,p.A254V消除了钙敏感性。p.G219C和p.G465R可能在质膜上部分不活跃。总之,大多数致病突变体部分或全部积聚在细胞内区室中,
更新日期:2020-03-26
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