The biosynthesis of mannolipids and mannose-containing complex glycans by the retina

Large-scale incubations were carried out with homogenates of the retinas of the 15--16-day-old chick embryo in the presence of GDP[U-14C] mannose, from which there were isolated mannolipid (Lipid I), oligosaccharide-lipids (Lipid II), and glycoprotein (residue). These incubations were performed in the presence of endogenous acceptors as well as dolichyl phosphate. [14C] Mannolipid I was subjected to chromatography on DEAE cellulose and silicic acid. The response to these, as well as TLC, enzymatic, and chemical treatments, were consistent with the product being dolichyl phosphomannose. [14C] Lipid II was purified by DEAE cellulose chromatography and gel filtration on LH-20. Responses to these treatments, as well as TLC and paper chromatography, were consistent with this product being of the class of the oligosaccharide-pyrophosphate-lipids. The residue remaining after removal of the lipids was shown to contain glycoproteins by conversion of high-molecular-weight radioactive material to low-molecular-weight [14C] mannose-containing glycopeptides by the action of pronase. These reactions and their products are consistent with there being in the retina, the pathway for glycoprotein synthesis involving the participation of the lipid-activated carbohydrates. When the incubations were performed in the presence of ATP or ADP there was a decrease in the labeling of Lipid I, accompanied by an increase in the labeling of Lipid II and glycoprotein. When incubated in the presence of dolichyl phosphate and detergent, however, the stimulatory effect of ATP did not occur. The effect on these activities of a variety of other nucleotide phosphates was also examined.