Topology of CNS myelin proteolipid protein: evidence for the nonenzymatic glycosylation of extracytoplasmic domains in normal and diabetic animals.
Myelin proteolipid protein (PLP), the main integral membrane protein in the central nervous system myelin, was labeled at the extracytoplasmic domains with the membrane impermeant reagents pyridoxal 5'-phosphate and tritiated borohydride. Lysine-217, located in the fourth hydrophilic domain of PLP, was found to be the major labeled residue, which defined this domain to be extracytoplasmic in agreement with our previously proposed topological model. The remarkably high reactivity in vitro of this residue as compared to all other lysines in PLP led us to investigate the possible modification of PLP in vivo by other carbonyl compounds. We demonstrate that PLP is the most highly nonenzymatically glycosylated membrane protein in murine and bovine brain. The degree of modification increases significantly under hyperglycemic conditions, as studied in diabetic mice. The majority of the glycosylation sites are also located at extracytoplasmic domains. The degree of nonenzymatic glycosylation of PLP may be related to late diabetic complications affecting the central nervous system.
Weimbs
T
T
Institute of Biochemistry, Medical Faculty, University of Cologne, Germany.
Stoffel
W
W
D000595:Amino Acid Sequence; D000818:Animals; D001665:Binding Sites; D001894:Borohydrides / Q000378:metabolism; D001921:Brain / Q000378:metabolism*; D002417:Cattle; D003593:Cytoplasm / Q000378:metabolism; D003921:Diabetes Mellitus, Experimental / Q000235:genetics / Q000378:metabolism*; D006031:Glycosylation; D066298:In Vitro Techniques; D008239:Lysine / Q000737:chemistry; D051379:Mice; D008810:Mice, Inbred C57BL; D008817:Mice, Mutant Strains; D008958:Models, Molecular; D008969:Molecular Sequence Data; D015394:Molecular Structure; D009185:Myelin Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D018991:Myelin Proteolipid Protein; D010084:Oxidation-Reduction; D011732:Pyridoxal Phosphate / Q000378:metabolism
D001894:Borohydrides; D009185:Myelin Proteins; D018991:Myelin Proteolipid Protein; D011732:Pyridoxal Phosphate; C025364:sodium borohydride; D008239:Lysine