Use of antibodies against dodecylsulfate-denatured heavy meromyosins to probe structural differences between muscular myosin isoenzymes.
Heavy meromyosin, a tryptic myosin fragment, was purified from rabbit fast twitch muscles and rat cardiac ventricles. Both types of heavy meromyosin were denatured by sodium dodecylsulfate and used to immunize guinea-pigs after chromatography on Sephadex G-10 to remove excess dodecylsulfate. Micro-complement fixation analysis showed that the antisera were specific to a denatured configuration of heavy meromyosin and myosin, and hardly recognized the native proteins. Cross-reactions performed with both rabbit skeletal and rat cardiac antisera indicated that the antigenic structures of denatured myosins varied according both to species (man, rabbit, rat or mouse), and to muscle-type (red skeletal slow twitch, while skeletal fast twitch, cardiac atria or cardiac ventricles). Denatured heavy meromyosin chromatography on Sephadex G-200 in the presence of 0.1% sodium dodecylsulfate enabled separation of several polypeptides groups. Of these, a polypeptide of Mr 29000 was the most reactive and exhibited the same immunological specificities as the whole myosin molecule. The use of antibodies against denatured heavy meromyosin in conjunction with micro-complement fixation therefore provides a discriminant means, not only for estimating the structural relationship between several myosin isoenzymes, but also for localizing constant and variable regions in the heavy chains of these isoenzymes.
D000818:Animals; D000906:Antibodies; D003429:Cross Reactions; D000939:Epitopes; D007106:Immune Sera; D007118:Immunoassay; D007527:Isoenzymes / Q000276:immunology; D008970:Molecular Weight; D009132:Muscles / Q000201:enzymology*; D009206:Myocardium / Q000201:enzymology; D015879:Myosin Subfragments / Q000276:immunology; D009218:Myosins / Q000276:immunology; D011817:Rabbits; D012967:Sodium Dodecyl Sulfate; D014357:Trypsin
D000906:Antibodies; D000939:Epitopes; D007106:Immune Sera; D007527:Isoenzymes; D015879:Myosin Subfragments; D012967:Sodium Dodecyl Sulfate; D014357:Trypsin; D009218:Myosins