m-Calpain requires DNA for activity on nuclear proteins at low calcium concentrations.
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Key Findings
Abstract
m-Calpain (calpain II, m-CANP), which normally requires millimolar Ca2+ for activity in vitro, was capable of proteolyzing a number of matrix proteins in isolated rat liver nuclei at Ca2+ concentrations as low as 3 microM (Mellgren, R. L. (1991) J. Biol. Chem. 266, 13920-13924). Treatment of nuclei with deoxyribonuclease I eliminated the activity of m-calpain at low Ca2+ concentrations, while ribonuclease A and phospholipase C had no effect. Addition of DNA to DNase-treated nuclei restored m-calpain activity at low Ca2+. RNA had little if any effect. Eukaryotic and prokaryotic DNA were equally effective, and synthetic polydeoxyribonucleotides were also activators. m-Calpain did not bind to a DNA-cellulose column in the presence of 200 microM Ca2+, and m-calpain preincubated in the presence of DNA and 200 microM Ca2+ was not activated at low Ca2+ concentrations following removal of the DNA. DNA did not alter the Ca2+ requirement for m-calpain-catalyzed cleavage of casein. These results demonstrate that the Ca2+ requirement for proteolysis of nuclear matrix proteins by m-calpain can be dramatically decreased in the presence of DNA. Activation did not seem to be a result of DNA binding directly to calpain but appeared to require interaction of DNA, calpain, and calpain substrates in the nuclear matrix.
Affiliation
Department of Pharmacology and Therapeutics, Medical College of Ohio, Toledo 43699.
External References
- PubMed ID:
- 8416968
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