极长链酰基辅酶A脱氢酶缺乏症ACADVL基因新发突变导致线粒体功能障碍（英文）

发布时间：2022-07-27 19:30

　　目的:鉴定并阐述6个ACADVL新发突变的对细胞和分子功能的影响,为后期分子诊断和临床病例管理提供科学依据。创新点:本文从分子功能和结构的角度阐述6个新位点对蛋白和细胞功能的不同影响。方法:通过体外构建突变载体建立各突变的过表达细胞系,检测各细胞系脂肪酸代谢能力、线粒体呼吸链功能、线粒体产三磷酸腺苷（ATP）能力、活性氧（ROS）和凋亡水平。同时检测了新发突变对蛋白二聚体稳定性的影响并用分子动力学模拟了突变蛋白构像的改变。结论:突变（c.541C>T、c.863T>G、c.895A>G、c.1238T>C、c.1276G>A和c.1505T>A）过表达细胞系均表现出脂肪酸代谢障碍,线粒体呼吸链功能障碍,ATP产生水平下降和线粒体ROS水平上升。在无糖条件下,各细胞系的凋亡水平上升。各突变影响了极长链酰基辅酶A脱氢酶（VLCAD）蛋白二聚体的稳定性,分子动力学模拟预测每个突变均会导致VLCAD蛋白结构的发生重要构象变化。 

【文章页数】：17 页

【文章目录】：
1 Introduction
2 Materials and methods
    2.1 Patients and novel variants
    2.2 Vector construction and cell culture
    2.3 Mitochondrial respiration assay
    2.4 FAO of cellular respiration and FAO flow cytometry
    2.5 ATP and mitochondrial superoxide measurements
    2.6 Western blotting
    2.7 Apoptosis assay
    2.8 Blue native PAGE
    2.9 Molecular dynamics simulation analyses
    2.1 0 Statistical analysis
3 Results
    3.1 Influence of variants on FAO capacity
    3.2 Influences of variants on mitochondrial res-piratory function,ATP production,and ROS
    3.3 Cells containing mutant VLCAD prone to apoptosis without glucose
    3.4 Influences of variants on the stability of VLCAD dimer and its MD simulation
4 Discussion
5 Conclusions
Contributors
Compliance with ethics guidelines
List of electronic supplementary materials



本文编号：3666043