Glycogen debranching enzyme: purification, antibody characterization, and immunoblot analyses of type III glycogen storage disease.

摘要: Type III glycogen storage disease is caused by a deficiency of debranching-enzyme activity. Many patients with this have both liver and muscle involvement, whereas others only involvement without clinical or laboratory evidence myopathy. To improve our understanding the molecular basis disease, debranching enzyme was purified 238-fold from porcine skeletal muscle. In sodium dodecyl sulfate-polyacrylamide gel electrophoresis gave single band relative weight 160,000 that migrated to same position as rabbit-muscle enzyme. Antiserum against prepared in rabbit. The antiserum reacted precipitin line demonstrated reaction having complete identity those present crude extracts. Incubation inhibited almost all activity, such treatment preimmune serum had little effect. also activity extracts pigs humans extent observed Immunoblot analysis probed anti-porcine-muscle showed can detect human liver. bands detected samples were size one Five six other types subjected immunoblot analysis. Although anti-porcine specific (Types I, II, IX), no cross-reactive material any disease. These data indicate (1) immunochemical similarity (2) due absence debrancher protein we studied.