Abstract
32P and 18O Exchange Reactions in Soluble Fractions Prepared from Rat Liver Mitochondria
Highlights
The Pi-Hz”0 exchange of i&act mitochondria was not inhibited by KF (Table VII). These findings suggest that the Pi-H2180 exchange activity of the soluble systems derived from mitochondria is unrelated to the Pi-H2180 exchange activity associated with respiratory chain-coupled phosphorylation in intact mitochondria
It has been shown that “Pi-ATP exchange activity of various soluble preparations derived from well washed mitochondria is greatly increased by addition of either glycolytic substrates or glyceraldehyde-3-P dehydrogenase and 3-P-glyceric P-kinase
It is likely that the effects of addition of glycolytic substrates indicate the presence of aldolase, triose isomerase, glyceraldehyde-3-P dehydrogenase, and 3-P-glyceric P-kinase activities in the soluble preparations; and the effects noted on addition of the last two enzymes mentioned are ascribable to the presence of PGA, glyceraldehyde-3-P, or a chemical moiety which can lead to their formation
Summary
Tissue PreparationsAll of the preparations described below were done at 0”.Standard Preparation of Mitochondria-Rat liver mitochondria were prepared as previously described [11] from albino male rats (200 to 250 g) purchased from the Badger Research Corporation, Madison, Wisconsin.Supernatant Fraction (105,000 x g) from Freeze-Thaw Treatment of Mitochondria (Fraction SF-FTM)i-This fraction was prepared essentially as described by Peter et cd. [12] from rat liver mitochondria.The mitochondria were isolated as in the preparation of standard mitochondria, except that 0.25 MNo RG-5606 from the Division of Research Grants of the National Institutes of Health, Public Health Service, Bethesda. All of the preparations described below were done at 0”. Standard Preparation of Mitochondria-Rat liver mitochondria were prepared as previously described [11] from albino male rats (200 to 250 g) purchased from the Badger Research Corporation, Madison, Wisconsin. Supernatant Fraction (105,000 x g) from Freeze-Thaw Treatment of Mitochondria (Fraction SF-FTM)i-This fraction was prepared essentially as described by Peter et cd. [12] from rat liver mitochondria. The mitochondria were isolated as in the preparation of standard mitochondria, except that 0.25 M. 105.000 X D supernatant fraction from freeze-thaw treatment of mitochondria; Fraction. SF-H, 105,000 X g supernatant fraction from whole liver homogenate; Fraction SF-TX1OOM,V, 105,000 X g supernatant fraction from Triton. X-100 treatment of mitochondria; PGA, 3-phosphoglyceric acid; DHAP, dihydroxyacetone phosphate
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