A simple and rapid method for the isolation of a large quantity of cytochrome c oxidase from bovine heart mitochondria was developed, based on selective solubilization of mitochondrial protein with first Triton and then lauryl maltoside. Gel filtration shows that the lauryl maltoside-solubilized oxidase preparation is in a hydrodynamically homogeneous state with a Stokes radius of 7.5 +/- 0.2 nm. It contains 8.0 mumol of haem (with an a/a3 ratio of 1)/g of protein. The catalytic constant (maximum turnover number) with respect to cytochrome c approaches 600 S-1. After further purification of the solubilized enzyme on a sucrose-gradient centrifugation, the purified enzyme has a haem content of 10.3 mumol/g of protein and eight major polypeptide bands shown on SDS/polyacrylamide-gel electrophoresis.
Research Article| March 01 1987
A new procedure for the purification of monodisperse highly active cytochrome c oxidase from bovine heart
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Biochem J (1987) 242 (2): 417-423.
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Y Li, A Naqui, T G Frey, B Chance; A new procedure for the purification of monodisperse highly active cytochrome c oxidase from bovine heart. Biochem J 1 March 1987; 242 (2): 417–423. doi: https://doi.org/10.1042/bj2420417
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