Amino-terminal domain stability mediates apolipoprotein E aggregation into neurotoxic fibrils.
The three isoforms of apolipoprotein (apo) E are strongly associated with different risks for Alzheimer's disease: apoE4>apoE3>apoE2. Here, we show at physiological salt concentrations and pH that native tetramers of apoE form soluble aggregates in vitro that bind the amyloid dyes thioflavin T and Congo red. However, unlike classic amyloid fibrils, the aggregates adopt an irregular protofilament-like morphology and are seemingly highly alpha-helical. The aggregates formed at substantially different rates (apoE4>apoE3>apoE2) and were significantly more toxic to cultured neuronal cells than the tetramer. Since the three isoforms have large differences in conformational stability that can influence aggregation and amyloid pathways, we tested the effects of mutations that increased or decreased stability. Decreasing the conformational stability of the amino-terminal domain of apoE increased aggregation rates and vice versa. Our findings provide a new perspective for an isoform-specific pathogenic role for apoE aggregation in which differences in the conformational stability of the amino-terminal domain mediate neurodegeneration.
Hatters
Danny M
DM
Gladstone Institute of Neurological Disease, 1650 Owens Street, San Francisco, CA 94158, USA.
Zhong
Ning
N
Rutenber
Earl
E
Weisgraber
Karl H
KH
D000818:Animals; D053329:Apolipoprotein E2; D053318:Apolipoprotein E3; D053327:Apolipoprotein E4; D001057:Apolipoproteins E / Q000737:chemistry* / Q000378:metabolism / Q000633:toxicity* / Q000648:ultrastructure; D052160:Benzothiazoles; D002478:Cells, Cultured; D003224:Congo Red; D051379:Mice; D008954:Models, Biological; D009474:Neurons / Q000187:drug effects* / Q000473:pathology; D020033:Protein Isoforms / Q000737:chemistry / Q000378:metabolism / Q000633:toxicity / Q000648:ultrastructure; D020836:Protein Structure, Quaternary; D017434:Protein Structure, Tertiary; D013329:Structure-Activity Relationship; D013816:Thermodynamics; D013844:Thiazoles
D053329:Apolipoprotein E2; D053318:Apolipoprotein E3; D053327:Apolipoprotein E4; D001057:Apolipoproteins E; D052160:Benzothiazoles; D020033:Protein Isoforms; D013844:Thiazoles; C009462:thioflavin T; D003224:Congo Red