Kinetics of formation of GlcNAc-GlcNAc-P-P-dolichol by microsomes from the retina of the embryonic chick.

Little quantitative information is available concerning individual reactions of the dolichol pathway. We have investigated the kinetics of the GlcNAc-transferase that catalyzes the biosynthesis of GlcNAc-GlcNAc-P-P-dolichol using chemically synthesized GlcNAc-P-P-dolichol as the substrate. Using microsomal preparations from the retina of the embryonic chick as enzyme source, optimal incubation conditions of pH, metal ion and detergent concentrations were established, after which apparent kinetic constants (Km and Vmax) were determined under initial rate conditions for GlcNAc-P-P-dolichol and UDP-GlcNAc. These studies provide the first quantitative description of the kinetics of this reaction.