Use of 2-D DIGE analysis reveals altered phosphorylation in a tropomyosin mutant (Glu54Lys) linked to dilated cardiomyopathy.
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.
Warren
Chad M
CM
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.
Arteaga
Grace M
GM
Rajan
Sudarsan
S
Ahmed
Rafeeq P H
RP
Wieczorek
David F
DF
Solaro
R John
RJ
D019943:Amino Acid Substitution / Q000235:genetics*; D000818:Animals; D002311:Cardiomyopathy, Dilated / Q000235:genetics* / Q000378:metabolism*; D015180:Electrophoresis, Gel, Two-Dimensional; D018698:Glutamic Acid / Q000235:genetics; D015151:Immunoblotting; D008239:Lysine / Q000235:genetics; D051379:Mice; D008822:Mice, Transgenic; D010766:Phosphorylation; D017354:Point Mutation; D014335:Tropomyosin / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*
D014335:Tropomyosin; D018698:Glutamic Acid; D008239:Lysine