Abstract
α-Connectin (also called titin 1) has been isolated from rabbit back muscle. Myofibrils were well washed with 5mM NaHCO3 and then extracted with 0.2M sodium phosphate, pH 7.0. The extract was dialyzed against 0.1M potassium phosphate, pH 7.0, to sediment myosin. The supernatant, adjusted to 0.18M potassium phosphate, pH 7.0, and 4M urea, was subjected to DEAE Toyopearl column chromatography.β-Connectin was eluted in the flow-through fraction and α-connectin was eluted at around 0.1M NaCl, when a 0 to 0.25M NaCl gradient was applied. The separated α-connectin was dialyzed against 0.2M potassium phosphate, pH 7.0. The resultant α-connectin showed the same mobility as that in an SDS extract of rabbit back muscle on SDS gel electrophoresis using 1.8% polyacrylamide gels. A monoclonal antibody against chicken breast muscle β-connectin reacted with the α-connectin isolated from rabbit back muscle.
