Cardiac muscle protein analysis by high-resolution and microscale two-dimensional gel electrophoresis.
An improved method of two-dimensional gel electrophoresis of small muscle samples is described. Isoelectric focusing of cardiac whole muscle homogenate in agarose gels containing urea and detergent has a markedly increased resolution. Equilibration of the first-dimensional gels with detergent before application to the second-dimensional gels is unnecessary in this system. By applying this method to rat cardiac whole muscle, high-molecular weight proteins, such as myosin heavy chains, are focused on the first-dimensional gels and, in addition, minor components are resolved on the second-dimensional gels, without loss during equilibration with detergent. The two-dimensional electrophoretic patterns of rat cardiac whole muscle obtained with this method reveal numerous clearly separated spots. By analyzing the two-dimensional electrophoretic patterns of rat cardiac whole muscle and various rat cardiac fractions, and by staining the calcium-binding proteins with "Stains-all", we identified some cardiac muscle components, such as myosin heavy and light chains, actin, tropomyosin, and troponin C, but additional work is required to identify the remaining spots. The two-dimensional electrophoretic system described here makes possible the effective resolution of whole cardiac muscle homogenate from small samples, and looks promising as an aid to muscle research.
Tanaka
T
T
The Third Division, Department of Internal Medicine, Osaka Medical College, Japan.
Kawamura
K
K
D000818:Animals; D015180:Electrophoresis, Gel, Two-Dimensional; D004591:Electrophoresis, Polyacrylamide Gel; D007525:Isoelectric Focusing; D008297:Male; D009124:Muscle Proteins / Q000032:analysis*; D009206:Myocardium / Q000032:analysis*; D051381:Rats; D011919:Rats, Inbred Strains; D013194:Staining and Labeling
D009124:Muscle Proteins