Apolipoprotein E structural requirements for the formation of SDS-stable complexes with beta-amyloid-(1-40): the role of salt bridges.
Of the three major isoforms of human apolipoprotein E (apoE), apoE4 is a risk factor for the development of Alzheimer's disease. Among possible neurologically relevant differences in the properties of apoE3 and apoE4 is the fact that apoE3 forms an SDS-stable complex with beta-amyloid-(1-40) (Abeta40) with greater avidity than does apoE4. This interaction may sequester potentially toxic species of Abeta or facilitate clearance. To understand more about this difference, we examined whether differences in salt bridges between apoE domains influence the capacity of apoE isoforms to form complexes with Abeta. In apoE3 there is a salt bridge between Arg-61 and Asp-65, while in apoE4 there are salt bridges between Arg-61 and Glu-255, and Arg-112 and Glu-109. Mutation of position 112, which is Cys in apoE3 and Arg in apoE4, to Ala or Lys abolished complex formation, while mutant apoE with Ser at this position retained the capacity to form complex. Substituting Ala for Glu-109 had no effect on the ability of either apoE4 or apoE3 to form complexes. On the other hand, substitution of Thr for Arg-61 in apoE3 abolished, and truncation of apoE3 at position 201 substantially lowered, but did not abolish, complex formation. Neither of these mutations within apoE4 had any affect on its complex formation with Abeta. These results suggest that the nature of the cysteine residue in apoE3 and interactions between the N-terminal and C-terminal domains of human apoE are important for the ability of apoE3 to form an SDS-stable complex with Abeta40.
Bentley
Nicholas M
NM
Department of Pathology, University of Chicago, 5841 S. Maryland Avenue, MC1089, Chicago, IL 60637-1470, U.S.A.
Ladu
Mary Jo
MJ
Rajan
Chandrika
C
Getz
Godfrey S
GS
Reardon
Catherine A
CA
D000544:Alzheimer Disease / Q000378:metabolism; D016229:Amyloid beta-Peptides / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D001057:Apolipoproteins E / Q000737:chemistry* / Q000378:metabolism*; D001120:Arginine / Q000737:chemistry; D003545:Cysteine / Q000737:chemistry; D018076:DNA, Complementary / Q000378:metabolism; D006801:Humans; D046911:Macromolecular Substances; D016297:Mutagenesis, Site-Directed; D010446:Peptide Fragments / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D011485:Protein Binding; D020033:Protein Isoforms; D017434:Protein Structure, Tertiary; D012492:Salts / Q000737:chemistry; D012967:Sodium Dodecyl Sulfate / Q000737:chemistry*
D016229:Amyloid beta-Peptides; D001057:Apolipoproteins E; D018076:DNA, Complementary; D046911:Macromolecular Substances; D010446:Peptide Fragments; D020033:Protein Isoforms; D012492:Salts; C078592:amyloid beta-protein (1-40); D012967:Sodium Dodecyl Sulfate; D001120:Arginine; D003545:Cysteine