The role of glycosylation in synthesis and secretion of beta-amyloid precursor protein by Chinese hamster ovary cells.
Alzheimer's disease is characterized by beta-amyloid deposition in the brain. This peptide is derived by proteolytic cleavage from beta-amyloid precursor protein (APP), a highly glycosylated membrane glycoprotein containing both N- and O-glycans. There are three major isoforms of APP, which are derived by alternative splicing and contain 695, 751, or 770 amino acids. Since glycosylation can affect many properties of glycoproteins, we studied the role of N- and O-glycosylation in the synthesis and secretion of APP. APP expression was examined in untransfected wild-type, Lec-8 mutant, and ldlD mutant Chinese hamster ovary (CHO) cells and in analogous clonal cell lines expressing either the transfected human wild-type 695-amino-acid form of APP (APP695-WT) or a form mutated to delete N-glycosylation sites (APP695-XX). These studies showed that maturation of APP in CHO cells is accompanied by the addition of multiple short O-glycans with the following structures: Neu5Acalpha2-3Galbeta1-3GalNAc, Neu5Acalpha2-3Galbeta1-3[Neu5Acalpha2-6]GalNAc, and GalNAc. Using glycosylation-defective mutant CHO cell lines and soluble inhibitors of glycosylation, we found that APP secretion was diminished when core N-glycosylation or N-glycan processing was blocked. Surprisingly, similar results were found when synthesis and secretion of either APP695-WT or APP695-XX were analyzed. These results indicate that defective N-glycosylation of other cellular proteins, but not of APP itself, affects the metabolism of APP. Interestingly, inhibition of O-glycosylation did not affect the biosynthesis or secretion of APP. The results of these studies may shed some light on the role that protein glycosylation may play in the pathogenesis of Alzheimer's disease.
Påhlsson
P
P
Department of Pathology and Laboratory Medicine, University of Pennsylvania, Philadelphia, Pennsylvania, 19104, USA.
Spitalnik
S L
SL
D016564:Amyloid beta-Protein Precursor / Q000378:metabolism*; D000818:Animals; D016466:CHO Cells; D006224:Cricetinae; D004791:Enzyme Inhibitors / Q000494:pharmacology; D006023:Glycoproteins / Q000378:metabolism*; D006031:Glycosylation; D007212:Indolizines / Q000494:pharmacology; D007700:Kinetics; D011499:Protein Processing, Post-Translational; D011994:Recombinant Proteins / Q000378:metabolism; D014162:Transfection; D014415:Tunicamycin / Q000494:pharmacology; D054836:Indolizidines
D016564:Amyloid beta-Protein Precursor; D004791:Enzyme Inhibitors; D006023:Glycoproteins; D007212:Indolizines; D011994:Recombinant Proteins; D014415:Tunicamycin; C037806:castanospermine; D054836:Indolizidines