Определение активности митохондриальной убихинол-цитохром с-оксидоредуктазы тромбоцитов в микропробах

V.T. Cheshchevik; N.G. Cheshchevik

Authors

V.T. Cheshchevik Polessky State University, Pinsk, Republic of Belarus
N.G. Cheshchevik Polessky State University, Pinsk, Republic of Belarus

Keywords:

platelets, mitochondria, ubiquinol-cytochrome c oxidoreductase, complex III, citrate synthase, cytochrome bc1 complex, electron transport chain

Abstract

The spectrophotometric method for determining the activity of ubiquinol-cytochrome c oxidoreductase (complex III) in mitochondria is one of the most widely used approaches. When measuring the activity of complex III in platelets, it is most practical to perform the measurements in microvolumes (100 µl) due to the limited availability of platelets from blood collection. The most suitable method for extracting the enzymatic activity of platelet mitochondrial complex III is a single freeze/thaw cycle of the platelet suspension. The optimal parameters for measuring the activity of platelet mitochondrial complex III are a temperature of 25°C, a protein amount in the sample of 80 µg, and concentrations of antimycin A at 90 µM, dodecylubiquinol at 100 µM, and cytochrome c at 75 µM, which ensure a stable enzyme reaction rate of complex III for 20 minutes in microvolumes. Normalization of complex III activity relative to platelet citrate synthase activity is only possible taking into account potential factors.

Author Biographies

V.T. Cheshchevik, Polessky State University, Pinsk, Republic of Belarus

PhD in Biol. Sc., Associate Professor,

Associate Professor of the Department of Biochemistry and Bioinformatics

N.G. Cheshchevik, Polessky State University, Pinsk, Republic of Belarus

Assistant Lecturer of the Department of Biochemistry and Bioinformatics

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