UDP-N-acetylglucosaminyl transferase (OGT) in brain tissue: temperature sensitivity and subcellular distribution of cytosolic and nuclear enzyme.
In brain tissue, UDP-N-acetylglucosaminyl transferase (OGT) is known to catalyze the addition of a single N-acetylglucosamine moiety (GlcNAc) onto two proteins linked to the etiology of neurodegenerative disease--beta-amyloid associated protein and tau. Hyperphosphorylation of tau appears to cause neurofibrillary tangles and cell death, and a functional relationship appears to exist between phosphorylation and glycosylation. Since a greater understanding of brain OGT may provide new insights into the pathogenesis of Alzheimer's disease, we examined the characteristics and subcellular distribution of OGT protein and OGT activity and its relationship to O-linked glycosylation. We found that cytosolic OGT activity is 10 times more abundant in brain tissue compared with muscle, adipose, heart, and liver tissue. Temperature studies demonstrated that cytosolic OGT activity was stable at 24 degrees C but was rapidly inactivated at 37 degrees C (T1/2 = 20 min). Proteases were probably not involved because OGT immunopurified from cytosol retained temperature sensitivity. Subcellular distribution studies showed abundant OGT protein in the nucleus that was enzymatically active. Nuclear OGT activity exhibited a high affinity for UDP-GlcNAc and a salt sensitivity that was similar to cytosolic OGT; however, nuclear OGT was not inactivated at 37 degrees C, as was the cytosolic enzyme. Two methods were used to measure O-linked glycoproteins in brain cytosol and nucleosol -[3H]galactose labeling and western blotting using antibodies against O-linked glycoproteins. Both methods revealed a greater abundance of O-linked glycoproteins in the nucleus compared to cytosol.
Okuyama
Ryo
R
Hexos, Inc., 18304 NE 153rd Street, Woodinville, WA 98072, USA. Hexos@comcast.net
Marshall
Stephen
S
D000273:Adipose Tissue / Q000201:enzymology; D000818:Animals; D000906:Antibodies / Q000378:metabolism; D001921:Brain / Q000201:enzymology*; D002467:Cell Nucleus / Q000737:chemistry / Q000201:enzymology*; D003600:Cytosol / Q000737:chemistry / Q000201:enzymology*; D004789:Enzyme Activation / Q000187:drug effects / Q000502:physiology; D004795:Enzyme Stability; D006023:Glycoproteins / Q000032:analysis; D008099:Liver / Q000201:enzymology; D018482:Muscle, Skeletal / Q000201:enzymology; D009206:Myocardium / Q000201:enzymology; D017351:N-Acetylglucosaminyltransferases / Q000737:chemistry* / Q000378:metabolism*; D009928:Organ Specificity; D010710:Phosphates / Q000494:pharmacology; D051381:Rats; D013347:Subcellular Fractions / Q000737:chemistry / Q000201:enzymology; D013696:Temperature; D014530:Uridine Diphosphate / Q000494:pharmacology; D014539:Uridine Diphosphate Sugars / Q000494:pharmacology
D000906:Antibodies; D006023:Glycoproteins; D010710:Phosphates; D014539:Uridine Diphosphate Sugars; C059191:uridine diphosphate N-acetylmannosamine; D014530:Uridine Diphosphate; D017351:N-Acetylglucosaminyltransferases; C063057:UDP-N-acetylglucosamine-peptide beta-N-acetylglucosaminyltransferase; C018279:sodium phosphate