Purification and properties of rat liver hydroxymethylglutaryl coenzyme A reductase phosphatases.

A procedure for the isolation and purification of two rat liver hydroxymethylglutaryl coenzyme A reductase phosphatases is described for the first time. Each of the preparations was obtained in two molecular forms of different molecular weights. The molecular weights of the holoenzymes were 480,000 and 310,000, respectively, while the molecular forms obtained after an ethanol treatment were in both cases 35,000. Several kinetic measurements were made which showed that the protein of Mr 35,000 was identical in both cases, irrespective of the holoenzymatic starting preparation used. The optimum pH of the three phosphatases ranged between 6.0 and 6.5. The Km of the phosphatases ranged between 6.5 and 19.5 nM when hydroxymethylglutaryl coenzyme A (HMG-CoA) reductase was the substrate. The three HMG-CoA reductase phosphatases, upon incubation, released 32P from 32P-labelled HMG-CoA reductase. This dephosphorylation also produces an activation of the HMG-CoA reductase activity.