Purification of cathepsin D from rabbit skeletal muscle and its action towards myofibrils.
Cathepsin D (EC 3.4.23.4) was purified from rabbit skeletal muscle using acetone-dried muscle powder as starting material. After the acetone-dried powder was extracted with 0.2 mM ATP, the extract was fractionated with acetone an subjected to DEAE-Sephadex A-50 and Sephadex G-100 column chromatography. Rechromatography on a Sephadex G-100 column resulted in a purified preparation. SDS-polyacrylamide gel electrophoresis of the purified enzyme showed one major band of 42,000 daltons and some bands of contaminants. Since gel filtration also indicated a value of 42,000 daltons for the enzyme, it was concluded that muscle cathepsin D has no subunit structure. The enzyme acted optimally towards myofibrils around pH3, resulting in the degradation of the myosin heavy chain and production of a 30,000-dalton component.
D000818:Animals; D002402:Cathepsin D; D002403:Cathepsins / Q000302:isolation & purification* / Q000378:metabolism; D008970:Molecular Weight; D009132:Muscles / Q000201:enzymology*; D009210:Myofibrils / Q000378:metabolism*; D011817:Rabbits
D002403:Cathepsins; D002402:Cathepsin D