Isolation and characterization of myosin heavy chain isozymes of the bovine conduction system.
To determine the characteristics of cardiac myosin in the conduction system, a pure Purkinje fiber preparation, consisting of atrioventricular nodes and the ventricular conduction system, was obtained from bovine hearts. Two types of myosin heavy chain isozymes, alpha-type and beta-type, were fractionated by affinity chromotography using monoclonal antibodies CMA19 and HMC50, which are specific for the alpha-type heavy chain and beta-type heavy chain, respectively. Competitive enzyme-linked immunosorbent assay demonstrated that the content of beta-type in the atrioventricular node (30-40%) was higher than that in atrial ordinary myocardium (10-20%) and that of the alpha-type was 30-40% in the ventricular conduction system, which was much higher than that in the ventricular ordinary myocardium (less than 10%). By one- and two-dimensional electrophoresis of the peptides produced by partial and complete digestion, the peptide compositions of alpha-type and beta-type in the conduction system were shown to be very similar to those of alpha-type and beta-type in ordinary myocardium, respectively. The CA2+-activated ATPase activity of myosin of the atrioventricular nodes was lower than that of ordinary atrial myosin (0.46 +/- 0.03 versus 0.58 +/- 0.02 mumol Pi/mg/min, mean +/- SEM, p less than 0.05) and in contrast, that of ventricular specialized myocardium was higher than that of myosin in the ventricular ordinary working myocardium (0.32 +/- 0.03 versus 0.22 +/- 0.01 mumol Pi/mg/min, p less than 0.05). This was in good agreement with the relative proportion of myosin isozymes.(ABSTRACT TRUNCATED AT 250 WORDS)
D000818:Animals; D000911:Antibodies, Monoclonal; D001283:Atrioventricular Node / Q000032:analysis; D000252:Calcium-Transporting ATPases / Q000378:metabolism; D002417:Cattle; D002846:Chromatography, Affinity; D004591:Electrophoresis, Polyacrylamide Gel; D004797:Enzyme-Linked Immunosorbent Assay; D006329:Heart Conduction System / Q000032:analysis*; D006352:Heart Ventricles / Q000032:analysis; D007158:Immunologic Techniques; D009206:Myocardium / Q000032:analysis; D009218:Myosins / Q000302:isolation & purification* / Q000378:metabolism; D010446:Peptide Fragments / Q000032:analysis; D011690:Purkinje Fibers / Q000032:analysis
D000911:Antibodies, Monoclonal; D010446:Peptide Fragments; D009218:Myosins; D000252:Calcium-Transporting ATPases