Abstract
Calpain (Ca 2+-dependent cysteine proteinase) is known to be a heterodimer, composed of one heavy (called 80K) and one light (called 30K) subunit. Calpain I, a low-Ca 2+-requiring form from porcine and human erythrocytes, and calpain II, a high-Ca 2+-requiring form from porcine kidney, were separated into their respective 80K and 30K subunits by high-performance liquid chromatography on a TSK-Gel G3000SW column in 1 m NaSCN. The isolated 80K subunits from porcine calpains I and II showed Ca 2+-dependent proteolysis, though much depressed and requiring higher Ca 2+ concentrations compared with the respective parent calpains. The optimal conditions were set for the reconstitution of a heterodimeric molecule from the once denatured and separated 80K and 30K subunits. In such reconstitution, the 30K subunits of calpains I and II were found to be functionally identical and interchangeable; either a homologous or heterologous 30K subunit lowered the Ca 2+ requirement of an 80K subunit significantly and enhanced the proteolytic activity. Subunit interchange could also be demonstrated between porcine and human calpains.
Published Version
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