Activation mechanism of calcium-activated neutral protease. Evidence for the existence of intramolecular and intermolecular autolyses |
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Authors: | M Inomata Y Kasai M Nakamura S Kawashima |
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Institution: | Department of Biochemistry, Tokyo Metropolitan Institute of Gerontology, Japan. |
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Abstract: | The activation mechanism through limited autolysis of a calcium-activated neutral protease (CANP) with a high sensitivity to calcium ions (microCANP) was analyzed. The rate of autolysis was dependent on microCANP concentration. The reaction was inhibited by high concentrations of digestible substrates but not by a nondigestible substrate. Incubation of microCANP inactivated by N-ethylmaleimide with a small amount of activated microCANP caused the degradation of the former in a manner similar to the autolysis of native microCANP. Immobilized microCANP bound to an anti-microCANP immunoglobulin G column autolyzed on addition of calcium ions. These results show that activation of microCANP through limited autolysis involves both intramolecular and intermolecular reactions. |
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