Kinetics of the coupled reaction catalysed by a fusion protein of yeast mitochondrial malate dehydrogenase and citrate synthase: Kinetics of a fusion protein of MDH and CS

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T1 - Kinetics of the coupled reaction catalysed by a fusion protein of yeast mitochondrial malate dehydrogenase and citrate synthase: Kinetics of a fusion protein of MDH and CS

AU - Pettersson, Henrik

AU - Olsson, Peter

AU - Bülow, Leif

AU - Pettersson, Gösta

PY - 2000

Y1 - 2000

N2 - The mechanistic implications of the kinetic behaviour of a fusion protein of mitochondrial malate dehydrogenase and citrate synthase have been reanalysed in view of predictions based on experimentally determined kinetic parameter values for the dehydrogenase and synthase activities of the protein. The results show that the time-course of citrate formation from malate in the coupled reaction catalysed by the fusion protein can be most satisfactorily accounted for in terms of a free-diffusion mechanism when consideration is taken to the inhibitory effects of NADH and oxaloacetate on the malate dehydrogenase activity. The effect of aspartate aminotransferase on the coupled reaction is likewise fully consistent with that expected for a free-diffusion mechanism. It is concluded that no tenable kinetic evidence is available to support the proposal that the fusion protein catalyses citrate formation from malate by a mechanism involving channelling of the intermediate oxaloacetate.

AB - The mechanistic implications of the kinetic behaviour of a fusion protein of mitochondrial malate dehydrogenase and citrate synthase have been reanalysed in view of predictions based on experimentally determined kinetic parameter values for the dehydrogenase and synthase activities of the protein. The results show that the time-course of citrate formation from malate in the coupled reaction catalysed by the fusion protein can be most satisfactorily accounted for in terms of a free-diffusion mechanism when consideration is taken to the inhibitory effects of NADH and oxaloacetate on the malate dehydrogenase activity. The effect of aspartate aminotransferase on the coupled reaction is likewise fully consistent with that expected for a free-diffusion mechanism. It is concluded that no tenable kinetic evidence is available to support the proposal that the fusion protein catalyses citrate formation from malate by a mechanism involving channelling of the intermediate oxaloacetate.

KW - fusion protein

KW - channelling

KW - malate dehydrogenase

KW - Citrate (si)-Synthase

KW - Oxaloacetic Acid

KW - Recombinant Fusion Proteins

KW - Saccharomyces cerevisiae

KW - NAD

KW - Models, Chemical

KW - Mitochondria

KW - Kinetics

U2 - 10.1046/j.1432-1327.2000.01558.x

DO - 10.1046/j.1432-1327.2000.01558.x

M3 - Article

C2 - 10931186

VL - 267

SP - 5041

EP - 5046

JO - European Journal of Biochemistry

JF - European Journal of Biochemistry

SN - 0014-2956

IS - 16

ER -