Native Complex Formation between Apolipoprotein E Isoforms and the Alzheimer's Disease Peptide Aβ

• Published in: Biochemistry (Easton) Vol. 35; no. 22; pp. 7123 - 7130
• Main Authors: Chan, Winnie, Fornwald, James, Brawner, Mary, Wetzel, Ronald
• Format: Journal Article
• Language: English
• Published: United States American Chemical Society 04.06.1996
• Subjects: 1-Propanol - pharmacology; Alzheimer Disease - metabolism; Amyloid beta-Peptides - chemistry; Amyloid beta-Peptides - metabolism; Apolipoproteins E - chemistry; Apolipoproteins E - genetics; Apolipoproteins E - metabolism; Chromatography, Gel; Electrophoresis, Polyacrylamide Gel; Escherichia coli; Escherichia coli - genetics; Humans; Immunoblotting; Isopropyl Thiogalactoside - pharmacology; Kinetics; Peptide Fragments - metabolism; Propanols; Protein Binding; Protein Conformation; Recombinant Proteins - isolation & purification; Recombinant Proteins - metabolism
• ISSN: 0006-2960; 1520-4995
• DOI: 10.1021/bi952852v

To explore whether the genetic linkage between apolipoprotein E (ApoE) alleles and susceptibility to Alzheimer's disease might be attributable to a direct molecular interaction between ApoE and the amyloid peptide Aβ, we have produced ApoE variants in Escherichia coli and studied their interactions with Aβ under native conditions. When incubated with Aβ at 20−40 μM concentrations, all three isoforms of ApoE (2, 3, and 4) readily form complexes with Aβ which can be isolated by gel filtration in native buffer. Freshly mixed ApoE and Aβ generate a complex that co-migrates in gel filtration with the main A 280 peak, which migrates identically to the ApoE tetramer alone. After several hours incubation, an additional, high molecular weight, soluble aggregate appears which also contains both ApoE and Aβ. Neither ApoE nor Aβ incubated by themselves produces high molecular weight aggregates under these conditions. Incubation of Aβ with control proteins bovine serum albumin and immunoglobulin generates negligable binding in the gel filtration assay. Similar results were obtained whether Aβ(1−40) or Aβ(1−42) was used, and plasma-derived ApoE gave similar results to E. coli-produced material. The data are consistent with a role for ApoE−Aβ interactions in modulating the development of AD. Since no major differences were observed in the behavior of the three ApoE isotypes, however, the molecular basis of the genetic trend between ApoE alleles and AD cannot be attributed to specific activity differences between the molecular forms of ApoE characterized in this study.