Differential Scanning Fluorimetry and Hydrogen Deuterium Exchange Mass Spectrometry to Monitor the Conformational Dynamics of NBD1 in Cystic Fibrosis.
Differential Scanning Fluorimetry and Hydrogen Deuterium Exchange Mass Spectrometry to Monitor the Conformational Dynamics of NBD1 in Cystic Fibrosis.
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差示扫描荧光法和氢氘交换质谱法监测囊性纤维化中 NBD1 的构象动力学。
DOI：
10.1007/978-1-4939-8820-4_4
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发表时间：
2019
期刊：
影响因子：
--
通讯作者：
Lukacs,GergelyL
中科院分区：
文献类型：
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作者：
Soya,Naoto;Roldan,Ariel;Lukacs,GergelyL
囊性纤维化 (CF) 是白种人中最常见的致命性常染色体隐性遗传疾病之一，预期寿命为 37-47 岁。 CF跨膜电导调节器（CFTR）是一种质膜离子通道，局限于上皮细胞的顶膜，并确保跨上皮水和溶质在多个器官中穿过分泌上皮细胞的运动。许多 CF 突变，包括核苷酸结合域 1 (NBD1) 中最常见的 F508 (ΔF508) 缺失，导致 CFTR 整体错误折叠和内质网 (ER) 的过早细胞内降解。为了更好地了解 NBD1 中 CF 引起的点突变引起的错误折叠机制（目前人们对此知之甚少），可以选择差示扫描荧光测定法 (DSF) 和氢氘交换质谱法 (HDX-MS)。这些已建立的方法可以通过监测主链酰胺 HDX 分别在全局和肽分辨率水平上测量 NBD1 的构象动力学，并将有助于评估 CF 突变和折叠校正器的作用机制，通过稳定突变 NBD1 来挽救 CFTR 折叠缺陷。
Cystic fibrosis (CF) is one of the most common, lethal autosomal recessive diseases in Caucasians with a life expectancy of 37–47 years. The CF transmembrane conductance regulator (CFTR) is a plasma membrane ion channel, confined to apical membrane of epithelia, and ensures transepithelial water and solute movement across secretory epithelia in several organs. Numerous CF mutations, including the most prevalent deletion of F508 (ΔF508) in the nucleotide binding domain 1 (NBD1) leads to CFTR global misfolding and premature intracellular degradation at the endoplasmic reticulum (ER). To better understand the misfolding mechanism caused by CF-causing point mutations in the NBD1, which is poorly understood, differential scanning fluorimetry (DSF) and hydrogen deuterium exchange coupled with mass spectrometry (HDX-MS) are the choice of techniques. These established methods can measure the conformational dynamics of the NBD1 globally and at peptide resolution level by monitoring backbone amide HDX, respectively, and will be instrumental to evaluate the mechanism of action of CF mutations and folding correctors that rescue CFTR folding defects via stabilizing the mutant NBD1.