Purification and some kinetic properties of rat liver ATP citrate lyase.
A new purification procedure for rat liver ATP citrate lyase is described. The method reproducibly gives homogenous undegraded enzyme. Steady-state kinetic analysis of ATP citrate lyase was complicated by the presence of ADP, a product of the reaction, in solutions of ATP. The kinetic patterns observed were dependent on whether ADP was removed by the assay system. When assays were performed with a method in which ADP was removed, the results showed that the enzyme obeys a double-displacement mechanism with a phosphoenzyme intermediate. This resolves a controversy between the results of previous kinetic studies and those of isotope-exchange and enzyme-labelling experiments.
D001275:ATP Citrate (pro-S)-Lyase / Q000302:isolation & purification / Q000378:metabolism*; D000244:Adenosine Diphosphate / Q000378:metabolism; D000255:Adenosine Triphosphate / Q000378:metabolism; D000818:Animals; D002951:Citrates / Q000378:metabolism; D019343:Citric Acid; D003065:Coenzyme A / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D005260:Female; D007700:Kinetics; D008099:Liver / Q000201:enzymology*; D008297:Male; D051381:Rats; D011919:Rats, Inbred Strains
D002951:Citrates; D019343:Citric Acid; D000244:Adenosine Diphosphate; D000255:Adenosine Triphosphate; D001275:ATP Citrate (pro-S)-Lyase; D003065:Coenzyme A