Dissociation and characterization of enzymes from a multienzyme complex involved in CO2 fixation.
A multienzyme complex from Euglena, molecular weight about 360,000, containing phosphoenolpyruvate carboxylase, malate dehydrogenase, and acetyl-coenzyme A carboxylase has been dissociated into active constituent enzymes. The respective molecular weights are 183,000, 67,000, and 127,000. The malate dehydrogenase contained in the complex is electrophoretically distinct from other malate dehydrogenase isozymes found in Euglena. The K-m for HCO3minus of the free and complexed acetyl-CoA carboxylase is 4.2-5.4 mM, and the substrate dependency for acetyl-CoA describes a sigmoidal relationship. The HCO3minus K-m for the free phosphoenolpyruvate carboxylase is 7.3-5.4 mM while that for the same enzyme contained in the complex is 0.7-1.3 mM. Both the free and complexed forms ofphosphoenolpyruvate carboxylase have a K-m for phosphoenolpyruvate of 0.9-1.7 mM. The latter enzyme in both the complex and free forms is stimulated by NADH, acetyl-CoA, and ATP. In the free phosphoenolpyruvate carboxylase, the stimulation passes through a maximum depending on effector concentration. The effect of NADH is to increase V-max while K-m values remain unmodified.
D000105:Acetyl Coenzyme A / Q000494:pharmacology; D000103:Acetyl-CoA Carboxylase / Q000378:metabolism*; D000255:Adenosine Triphosphate / Q000494:pharmacology; D000818:Animals; D001639:Bicarbonates / Q000494:pharmacology; D002245:Carbon Dioxide / Q000378:metabolism*; D005056:Euglena gracilis / Q000201:enzymology*; D007700:Kinetics; D008025:Ligases / Q000378:metabolism*; D008291:Malate Dehydrogenase / Q000378:metabolism*; D008970:Molecular Weight; D009097:Multienzyme Complexes / Q000032:analysis*; D009243:NAD / Q000494:pharmacology; D010729:Phosphoenolpyruvate Carboxykinase (GTP) / Q000378:metabolism*
D001639:Bicarbonates; D009097:Multienzyme Complexes; D009243:NAD; D002245:Carbon Dioxide; D000105:Acetyl Coenzyme A; D000255:Adenosine Triphosphate; D008291:Malate Dehydrogenase; D010729:Phosphoenolpyruvate Carboxykinase (GTP); D008025:Ligases; D000103:Acetyl-CoA Carboxylase