二维差异凝胶电泳（2-D DIGE）分析的应用揭示了与扩张型心肌病相关的原肌球蛋白突变体（Glu54Lys）中磷酸化的改变。
Use of 2-D DIGE analysis reveals altered phosphorylation in a tropomyosin mutant (Glu54Lys) linked to dilated cardiomyopathy.
作者信息
Warren Chad M, Arteaga Grace M, Rajan Sudarsan, Ahmed Rafeeq P H, Wieczorek David F, Solaro R John
机构信息
Department of Physiology and Biophysics, Center for Cardiovascular Research, College of Medicine, University of Illinois at Chicago, 835 E. Wolcott Avenue, Chicago, IL 60612, USA.
出版信息
Proteomics. 2008 Jan;8(1):100-5. doi: 10.1002/pmic.200700772.
Abstract
Current electrophoretic methods have not been optimized to fully separate post-translationally modified mutant forms of tropomyosin (Tm) from wild-type cardiac samples. We describe here a method employing a modified 2-D PAGE/2-D DIGE protocol, to fully separate native, mutant (E54K), and phosphorylated forms of Tm. Our data demonstrate the first evidence of a significant (approximately 40%) decrease in Tm phosphorylation in transgenic compared to non-transgenic mouse hearts, and indicate that altered phosphorylation may be a significant factor in the linkage of the E54K mutation to dilated cardiomyopathy.
摘要
目前的电泳方法尚未得到优化，无法从野生型心脏样本中完全分离翻译后修饰的原肌球蛋白（Tm）突变形式。我们在此描述一种采用改良的二维聚丙烯酰胺凝胶电泳/二维差异凝胶电泳方案的方法，以完全分离Tm的天然、突变（E54K）和磷酸化形式。我们的数据首次证明，与非转基因小鼠心脏相比，转基因小鼠心脏中Tm磷酸化显著降低（约40%），并表明磷酸化改变可能是E54K突变与扩张型心肌病关联的一个重要因素。
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