FIELD: medicine.
SUBSTANCE: method can be used for quantitative evaluation of superoxide in membrane preparations (microsomes, mitochondria) expressing cytochrome C reductase and cytochrome C oxydase activity. Besides its can be used in medical biochemistry to study the functions of superoxide in pathogenesis of neurodegenerative diseases. The method includes preparation of a measuring medium, mixing of a superoxide source and said measuring medium, addition to the prepared mixture of an oxidation substratum and mutant cytochrome with K27E/E69K/K72E/K86E/K87E/E90K or K8E/E62K/E69K7K72E/K86E/K87E, or K8E/K27E/E62K/E69K/K72E/K86E/K87E/E90K. It is followed with spectrophotometric measurement of cytochrome C derivative reduction rate and determination of superoxide generation rate.
EFFECT: method is simpler in comparison with common analogues and allows for high-accurate determination of superoxide generation rate.
1 tbl, 4 dwg, 1 ex
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