Cholesterol decreases secretion of the secreted form of amyloid precursor protein by interfering with glycosylation in the protein secretory pathway.
Cerebral deposits of beta-amyloid (betaA) are a major feature in Alzheimer's disease. betaA is derived from amyloid precursor protein (APP). APP is subject to N- and O-glycosylation and undergoes a series of proteolytic cleavages that lead to the release of betaA or of a non-amyloidogenic secreted form of APP (APPs). We used primary neuronal and glial cultures to investigate how cholesterol affects the production and secretion of APPs. Exposure to cholesterol for 2 h did not change the neuronal release of APPs; after 6 h APPs release was slightly lower, whereas 24 h of exposure decreased APPs in the medium by approx. 60%. The time courses were similar in astrocytes and microglia preparations. To verify whether the effect of cholesterol was a consequence of membrane rigidification we tested the activity of ganglioside GM1 and prion protein fragment PrP 106-126, which affect membrane fluidity similarly to cholesterol, on APPs secretion. Neither altered the production of APPs. APP mRNA and the total amount of APP in the cells were slightly decreased by cholesterol after 2 and 24 h respectively. Immunoblot analysis of APP associated with neuronal cells and astrocytes indicated that cholesterol progressively decreased the glycosylated forms of the protein; a similar tendency was noted in cells treated with brefeldin A and monensin, two substances that interfere with protein glycosylation. The cell-surface biotinylation method showed that in cholesterol-treated cells APP reached the plasma membrane. Our results indicate that cholesterol decreases the secretion of APPs by interfering with APP maturation and inhibiting glycosylation of the protein; although APP is inserted in the membrane it is not cleaved by alpha-secretase.
Galbete
J L
JL
Laboratory of Biology of Neurodegenerative Disorders, Istituto di Ricerche Farmacologiche 'Mario Negri', 20157 Milano, Italy.
Martin
T R
TR
Peressini
E
E
Modena
P
P
Bianchi
R
R
Forloni
G
G
D016564:Amyloid beta-Protein Precursor / Q000235:genetics* / Q000378:metabolism; D000818:Animals; D000831:Animals, Newborn; D001253:Astrocytes / Q000166:cytology / Q000187:drug effects / Q000502:physiology; D001921:Brain / Q000166:cytology / Q000502:physiology*; D002478:Cells, Cultured; D002784:Cholesterol / Q000494:pharmacology*; D005333:Fetus; D005677:G(M1) Ganglioside / Q000494:pharmacology; D005786:Gene Expression Regulation / Q000187:drug effects; D006031:Glycosylation / Q000187:drug effects; D007700:Kinetics; D008560:Membrane Fluidity / Q000187:drug effects / Q000502:physiology; D051379:Mice; D009457:Neuroglia / Q000166:cytology / Q000187:drug effects / Q000502:physiology*; D009474:Neurons / Q000166:cytology / Q000187:drug effects / Q000502:physiology*; D010446:Peptide Fragments / Q000494:pharmacology; D011328:Prions / Q000494:pharmacology; D051381:Rats; D013997:Time Factors; D014158:Transcription, Genetic / Q000187:drug effects
D016564:Amyloid beta-Protein Precursor; D010446:Peptide Fragments; D011328:Prions; D005677:G(M1) Ganglioside; D002784:Cholesterol