Plant Physiol. Journal of Pharmacology and Experimental Therapeutics
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Plant Physiology 64:150-153 (1979)
© 1979 American Society of Plant Biologists

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Articles

Separation of Mitochondria from Contaminating Subcellular Structures Utilizing Silica Sol Gradient Centrifugation 1

Christopher Jackson, Jane E. Dench, David O. Hall and Anthony L. Moore2

a Department of Plant Sciences, University of London King's College, 68 Half Moon Lane, London SE24 9JF United Kingdom

Discontinuous Percoll density gradients have been developed for the purification of mitochondria, permitting rapid separation under isosmotic and low viscosity conditions. Mitochondria from several etiolated tissues have been successfully separated from contaminating subcellular structures by this method. For potato tuber the ratio of washed to purified mitochondrial protein was 2.6, similar to the increase in specific activity of cytochrome c oxidase following separation. The purification of mitochondria from green leaf tissues on Percoll gradients has reduced chlorophyll contamination of spinach mitochondria from about 70 micrograms chlorophyll per milligram protein to approximately 8 micrograms chlorophyll per milligram protein.

The ratio of protein content of the washed mitochondria compared to that in the purified preparation was 7 for spinach and respiratory activity was retained. The physiological integrity and oxidative properties of washed and gradient mitochondria are compared.

2 Permanent address and to whom correspondence should be addressed: Department of Biochemistry, The University of Sussex, Falmer, Brighton, Sussex BN1 RH.

1 This work was supported by a grant from the Rank Prize Research Fund.




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