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Oxidative Phosphorylation in Ascites Tumor Mitochondria^*

Leonard A. Sauer, Arlene P. Martin and E. Stotz

( Department of Biochemistry, University of Rochester School of Medicine and Dentistry, Rochester, N.Y.)

A modified method for the cytochemical fractionation of an ascites tumor is described. The cells are pretreated in 10^-3 M EDTA in 0.25 M sucrose before homogenization. Particulate fractions are obtained by the usual centrifugation procedures.

The capacity of the isolated mitochondrial fraction to undergo oxidative phosphorylation was investigated. Mitochondria were found to be active with the substrates succinate, malate, and glutamate, and P/O ratios approaching the theoretical were obtained. The use of malonate in conjunction with glutamate oxidation was necessary to obtain maximal P/O ratios. No exogenous DPN was necessary for optimal oxygen uptake. Added DPN usually inhibited respiration and phosphorylation, especially the latter. A possible explanation for this effect is given.

^* This research has been partially supported by Grant No. H 1322, National Heart Institute, National Institutes of Health.

Medical Student Fellow, PHS Experimental Training Grant 2R-6. Present address: The Rockefeller Institute, New York 21, New York.

Received 12/21/61.