Presence of a pathway for biosynthesis of UDP-glucose in the Golgi apparatus of rat hepatocytes

Glucokinase, phosphoglucomutase and glucose-1-phosphate uridylyltransferase are the three enzymes involved in a microsomic pathway for the synthesis of UDP glucose. Evidence is given, in this paper, for the localization of these three enzymes in a Golgi-rich fraction of rat liver. This fraction is prepared, from smooth microsomes, by the means of a discontinuous four-step sucrose gradient. Three of the lighter fractions (d = 1.08-1.13) are enriched in the Golgi markers (galactosyltransferase, sialytransferase and thiamin pyrophosphatase), especially the one with density 1.13. The three enzymes we are interested in are enriched in the two upper hands (d 1.08-1.11), which display an activity for the biosynthesis of UDP-glucose from glucose equivalent to the one obtained in a crude microsomic preparation, and which are not contaminated by other subcellular components.