Department of Neurology, Yonsei University College of Medicine, Seoul, Republic of Korea.
Departments of Neurology, Mokdong Hospital, Ewha Womans University School of Medicine, Seoul, Republic of Korea.
J Clin Neurosci. 2020 Jan;71:186-190. doi: 10.1016/j.jocn.2019.08.068. Epub 2019 Aug 19.
Dysferlinopathy is an autosomal recessive disease caused by pathogenic variants in DYSF gene. We compared muscle protein extracts from dysferlinopathy patients and control subjects to identify new biomarkers of this myopathy. We reviewed the medical records from January 2002 to October 2016. Eight vastus lateralis muscle samples from five dysferlinopathy patients and three control subjects were selected. We separated proteins/peptides from all eight muscle protein extracts using two-dimensional electrophoresis (2DE). Data were acquired from liquid chromatography-mass spectrometry protein fragmentation patterns after comparing the spot volumes. Western blotting revealed total dysferlin loss in the dysferlinopathy patients but normal expression in the control subjects. 2DE indicated somewhat diverse protein constellations between the dysferlinopathy and control groups. Image analysis showed that 80 spots were differently expressed between two dysferlinopathy and one control samples. We selected 44 spots with consistently different volume between dysferlinopathy and control groups. Liquid chromatography-mass spectrometry indicated 26 differently expressed proteins. Western blotting revealed that creatine kinase M-type, carbonic anhydrase III (muscle specific) and desmin were significantly elevated in dysferlinopathy muscle. Additionally, four proteins (myosin light chain 1/3, skeletal muscle isoform; lamin A/C; ankyrin repeat domain 2; and eukaryotic translation initiation factor 5A-1) were inconsistently elevated in the dysferlinopathy samples. We confirmed the usefulness of the classic biomarker and have newly identified the altered expression of proteins in the skeletal muscles of dysferlinopathy patients.
肌营养不良蛋白病是一种由 DYSF 基因突变引起的常染色体隐性遗传病。我们比较了肌营养不良蛋白病患者和对照组的肌肉蛋白提取物，以鉴定这种肌病的新生物标志物。我们回顾了 2002 年 1 月至 2016 年 10 月的病历。从五例肌营养不良蛋白病患者和三例对照组中选择了 8 个股外侧肌样本。我们使用二维电泳（2DE）从所有 8 个肌肉蛋白提取物中分离蛋白质/肽。通过比较斑点体积，从液相色谱-质谱蛋白片段化模式中获取数据。Western blot 显示肌营养不良蛋白病患者的总肌营养不良蛋白缺失，但对照组正常表达。2DE 表明肌营养不良蛋白病组和对照组之间的蛋白质组成有些不同。图像分析显示，在两个肌营养不良蛋白病样本和一个对照组样本之间有 80 个斑点表达不同。我们选择了 44 个在肌营养不良蛋白病组和对照组之间体积差异始终存在的斑点。液相色谱-质谱分析显示 26 种差异表达蛋白。Western blot 显示肌营养不良蛋白病肌肉中的肌酸激酶 M 型、碳酸酐酶 III（肌肉特异性）和结蛋白显著升高。此外，在肌营养不良蛋白病样本中，有 4 种蛋白（肌球蛋白轻链 1/3、骨骼肌同工型；核纤层蛋白 A/C；锚蛋白重复结构域 2；真核翻译起始因子 5A-1）表达不一致升高。我们证实了经典生物标志物的有用性，并新发现了肌营养不良蛋白病患者骨骼肌中蛋白质表达的改变。
