Metabolic labeling of dolichol and dolichyl phosphate in isolate hepatocytes.

Abstract

Isolated hepatocytes from rat liver were incubated with [3H]mevalonate, and the labeling of polyprenols in the microsomal fraction was followed. After a 1-min incubation the alpha-unsaturated forms of polyprenyl-P2, -P, and polyprenol were mainly labeled and at this time point only 2, 8, and 17%, respectively, of the label was associated with the saturated forms. In the case of the free alcohol 2 h of incubation was required before all the labeling was recovered in the saturated form. After 1 min polyprenols and polyprenyl-P with 20 and 21 isoprene residues demonstrated much higher specific labeling than the shorter compounds, but after 5 min these differences were greatly reduced. In experiments utilizing short incubation times and chasing no evidence has been obtained that the phosphorylated form is a precursor of the free alcohol or vice versa, that the free alcohol is a precursor of the phosphorylated form. In human liver about 1% of the dolichol is present in the alpha-unsaturated form. These experiments suggest that: 1) the alpha-unsaturated form is the precursor of the alpha-saturated free alcohol, 2) dolichol does not necessarily arise directly from dephosphorylation of the phosphorylated form, and 3) the free alcohol is for the most part not phosphorylated under in vivo conditions in rat liver.