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Figure 1


Fig. 1. Interaction of AMPK {gamma}1 subunit with plectin. (A) Schematic representation showing the domain structure of plectin and the fragment (ple-R5-6; I4218-G4543; GenBank accession number AY480038) used as bait in the yeast two-hybrid screening (Osmanagic-Myers and Wiche, 2004). The actin- and IF-binding domains (ABD, IFBD), the rod and C-terminal repeat domains 1-6 of plectin are indicated. The isolated AMPK {gamma}1 subunit clone contained the entire coding sequence comprising four CBS domains. (B) Plectin was immunoprecipitated from lysates of wild-type (+/+) myotubes using anti-plectin antiserum. Lysates (Lys) and immunoprecipitates (IP) were analysed by immunoblotting using antibodies immunoreactive with AMPK {gamma}1 subunit and plectin. Note Co-IP of AMPK {gamma}1, and absence of signal in control sample (C) run without precipitating antibodies and in sample from plectin-deficient myocytes (-/-). (C) Immunoprecipitation of {alpha}1, {alpha}2, and {gamma}1 AMPK subunits from lysates of differentiated wild-type myocytes (myotubes) using indicated antibodies, and detection of cosedimenting {alpha}1 and {gamma}1 AMPK subunits by immunoblotting.