Purification and some characteristics of a Ca2+-activated proteolytic enzyme from beef cardiac muscle
- 1 April 1981
- journal article
- research article
- Published by Canadian Science Publishing in Canadian Journal of Biochemistry
- Vol. 59 (4) , 242-249
- https://doi.org/10.1139/o81-033
Abstract
A calcium-dependent neutral proteinase was purified from beef cardiac muscle. The crude extract prepared from cardiac muscle was subjected to acid precipitation and salt fractionation and then further purified by column chromatography on Sepharose 6B, DE-52, and Sephadex G-200 columns in succession. The final preparation showed an 11 300 fold increase in specific activity of the Ca2+-activated enzyme. Average enzyme protein yield was 2.4 μg/g fresh tissue. The enzyme was maximally active at pH 7.6 in the presence of 4 mM calcium. Proportionality of enzyme activity in partially purified preparations was retained when activity was measured at 25 °C using casein as the substrate. The rate of proteolysis by the purified enzyme was linear for 60 min under similar assay conditions. Fractionation of muscle homogenates showed that 70 to 73% of the total enzyme activity was present in the 24 000 × g and 30 000 × g supernatants. The enzyme was labile in aqueous solutions and storage at 4 °C and −20 °C resulted in considerable loss of activity, unless glycerol (50% v/v) was added to the solution.This publication has 6 references indexed in Scilit:
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