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. 1969 Sep;114(3):597–610. doi: 10.1042/bj1140597

The kinetic properties of citrate synthase from rat liver mitochondria

D Shepherd 1, P B Garland 1
PMCID: PMC1184933  PMID: 5820645

Abstract

1. Citrate synthase (EC 4.1.3.7) was purified 750-fold from rat liver. 2. Measurements of the Michaelis constants for the substrates of citrate synthase gave values of 16μm for acetyl-CoA and 2μm for oxaloacetate. Each value is independent of the concentration of the other substrate. 3. The inhibition of citrate synthase by ATP, ADP and AMP is competitive with respect to acetyl-CoA. With respect to oxaloacetate the inhibition by AMP is competitive, but the inhibition by ADP and ATP is mixed, being partially competitive. 4. At low concentrations of both substrates the inhibition by ATP is sigmoidal and a Hill plot exhibits a slope of 2·5. 5. The pH optimum of the enzyme is 8·7, and is not significantly affected by ATP. 6. Mg2+ inhibits citrate synthase slightly, but relieves the inhibition caused by ATP in a complex manner. 7. At constant total adenine nucleotide concentration made up of various proportions of ATP, ADP and AMP, the activity of citrate synthase is governed by the concentration of the sum of the energy-rich phosphate bonds of ADP and ATP. 8. The sedimentation coefficient of the enzyme, as measured by activity sedimentation, is 6·3s, equivalent to molecular weight 95000.

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Selected References

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