Oligomerization of amyloid beta-protein occurs during the isolation of lipid rafts.
Cholesterol- and glycosphingolipid-rich microdomains, called "lipid rafts," are suggested to initiate and promote the pathophysiology of Alzheimer's disease by serving as a platform for generation, aggregation, or degradation of amyloid-beta protein (Abeta). However, methods for biochemical isolation of these microdomains may produce artifacts. In this study, when synthetic Abeta1- 40 monomers were added to the brain fragment at a final concentration of 2.1 microM, followed by homogenization and isolation of lipid rafts by an established method, Abeta1- 40 accumulated as oligomers in the lipid raft fraction. However, in the absence of a brain homogenate, synthetic Abeta1- 40 did not accumulate in the lipid raft fraction. When fractionation was performed in the absence of synthetic Abeta1-40 and synthetic Abeta1-40 was incubated in an aliquot of each fraction, a marked oligomerization of Abeta1- 40 was observed in the lipid raft aliquot. These results indicate that exogenous Abeta associates with lipid rafts, and Abeta bound to rafts forms oligomers during the isolation of lipid rafts. In addition, endogenous Abeta1-40 in a Triton X-100-insoluble fraction of a brain homogenate of the Tg2576 transgenic mouse model of Alzheimer's disease formed oligomers when the fraction was incubated at 4 degrees C for 20 hr. Thus, one should be careful when one discusses the role of lipid rafts in amyloid precursor protein processing and in the generation, aggregation, and degradation of Abeta.
Yu
Wenxin
W
Department of Alzheimer's Disease Research, National Institute for Longevity Sciences, Aichi, Japan.
Zou
Kun
K
Gong
Jian-Sheng
JS
Ko
Mihee
M
Yanagisawa
Katsuhiko
K
Michikawa
Makoto
M
D000544:Alzheimer Disease / Q000473:pathology; D016229:Amyloid beta-Peptides / Q000737:chemistry*; D000818:Animals; D016477:Artifacts; D001923:Brain Chemistry; D004195:Disease Models, Animal; D004591:Electrophoresis, Polyacrylamide Gel; D008055:Lipids / Q000302:isolation & purification*; D021962:Membrane Microdomains / Q000737:chemistry*; D051379:Mice; D008822:Mice, Transgenic; D051381:Rats
D016229:Amyloid beta-Peptides; D008055:Lipids