Warren Chad M, Arteaga Grace M, Rajan Sudarsan, Ahmed Rafeeq P H, Wieczorek David F, Solaro R John
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.
Proteomics. 2008 Jan;8(1):100-5. doi: 10.1002/pmic.200700772.
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.
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。
