Isolation of the human ATP-creatine transphosphorylases (creatine phosphokinases) from tissues of patients with Duchenne muscular dystrophy.

Detailed procedures are described for the separation and isolation of the isoenzymes of ATP-creatine transphosphorylase (creatine phosphokinase) from autopsy samples of muscle and brain tissue of five terminal dystrophic males (x-linked, Duchenne). These direct isolation studies have led to the remarkable observation that three isoenzymes are present in the atrophied muscle tissue (characterized electrophoretically and immunologically as muscle type, hybrid type, and brain type, i:? order of decreasing amounts), but only one isoenzyme (brain type) is present in the dystrophic brain tissue. The muscle type was isolated in crystalline form, the other muscle isoenzymes (hybrid and brain type) were purified, the hybrid type was also purified from the heart tissue, and the brain type from the dystrophie brain was crystallized. In the normal male adult, however, only one isoenzyme appears to be present in the muscle (the muscle type) and only one isoenzyme (the brain type) in the brain. Also, studies on a stillborn and an ll-monthold male infant, revealed only the muscle type with traces of the hybrid type, and the absence of the brain type in the skeletal muscle. A sample obtained from the muscle of an llto 12-week-old female fetus also yielded the three-isoenzyme distribution pattern, characterized electrophoretically as muscle type, hybrid type, and brain type. Therefore, the isoenzymic distribution pattern of ATP-creatine transphosphorylase in the atrophied musculature of the terminal human progressive muscular dystrophic organism appears strikingly similar to that of the human fetal muscle.