Subcellular sites of enzymes catalyzing the phosphorylation-dephosphorylation of dolichol in the central nervous system.
The subcellular locations of several enzymes involved in dolichyl monophosphate (Dol-P) metabolism in brain have been investigated. Dolichol kinase is highly enriched in a heavy microsomal fraction from calf brain, while 71% of the Dol-P phosphatase activity was recovered with the light microsomes. Lower amounts of the phosphatase activity were also found in the heavy microsomal, mitochondrial-lysosomal, and synaptic plasma membrane fractions. Since the light microsomal fraction also contained substantial acetylcholinesterase activity, an axon plasma membrane marker, an axolemma-enriched fraction, was prepared from rat brain by a second procedure. A comparison with microsomal and mitochondrial-lysosomal fractions revealed that the axolemma-enriched fraction contained the highest specific activity of Dol-P phosphatase, indicating that the enzyme was present in the axon plasma membrane. The tunicamycin-sensitive UDP-N-acetylglucosamine:Dol-P N- acetylglucosaminylphosphotransferase , glucosyl- phosphoryldolichol (Glc-P-Dol) synthase, Glc-P-Dol:oligosaccharide glucosyltransferase, and the oligosaccharyltransferase were all found predominantly in the heavy microsomes. These results indicate that the enzymes responsible for the initiation and termination of biosynthesis, as well as the transfer of dolichol-linked oligosaccharides, reside in the rough endoplasmic reticulum (ER) of central nervous tissue. Evidence that at least some Dol-P molecules formed by dolichol kinase are accessible to multiple glycosyltransferases in the rough ER of brain is also presented.
D000818:Animals; D001665:Binding Sites; D001921:Brain / Q000201:enzymology*; D002384:Catalysis; D002417:Cattle; D004288:Dolichol Phosphates / Q000378:metabolism*; D005964:Glucosyltransferases / Q000302:isolation & purification; D006602:Hexosyltransferases; D008565:Membrane Proteins; D008861:Microsomes / Q000201:enzymology; D008928:Mitochondria / Q000201:enzymology; D010744:Phosphoric Monoester Hydrolases / Q000302:isolation & purification*; D010766:Phosphorylation; D010770:Phosphotransferases / Q000302:isolation & purification*; D017853:Phosphotransferases (Alcohol Group Acceptor); D011106:Polyisoprenyl Phosphates / Q000378:metabolism*; D051381:Rats; D013347:Subcellular Fractions / Q000201:enzymology*; D014166:Transferases / Q000302:isolation & purification
D004288:Dolichol Phosphates; D008565:Membrane Proteins; D011106:Polyisoprenyl Phosphates; D014166:Transferases; D005964:Glucosyltransferases; D006602:Hexosyltransferases; C025620:dolichyl-diphosphooligosaccharide - protein glycotransferase; D010770:Phosphotransferases; D017853:Phosphotransferases (Alcohol Group Acceptor); C020087:dolichol kinase; D010744:Phosphoric Monoester Hydrolases; C020088:dolichyl-phosphatase