title: Structural differences between apolipoprotein E3 and E4 as measured by (19)F NMR.
abstract: The apolipoprotein E family contains three major isoforms (ApoE4, E3, and E2) that are directly involved with lipoprotein metabolism and cholesterol transport. ApoE3 and apoE4 differ in only a single amino acid with an arginine in apoE4 changed to a cysteine at position 112 in apoE3. Yet only apoE4 is recognized as a risk factor for Alzheimer's disease. Here we used (19)F NMR to examine structural differences between apoE4 and apoE3 and the effect of the C-terminal domain on the N-terminal domain. After incorporation of 5-(19)F-tryptophan the 1D (19)F NMR spectra were compared for the N-terminal domain and for the full length proteins. The NMR spectra of the N-terminal region (residues 1-191) are reasonably well resolved while those of the full length wild-type proteins are broad and ill-defined suggesting considerable conformational heterogeneity. At least four of the seven tryptophan residues in the wild type protein appear to be solvent exposed. NMR spectra of the wild-type proteins were compared to apoE containing four mutations in the C-terminal region that gives rise to a monomeric form either of apoE3 under native conditions (Zhang et al., Biochemistry 2007; 46: 10722-10732) or apoE4 in the presence of 1 M urea. For either wild-type or mutant proteins the differences in tryptophan resonances in the N-terminal region of the protein suggest structural differences between apoE3 and apoE4. We conclude that these differences occur both as a consequence of the Arg158Cys mutation and as a consequence of the interaction with the C-terminal domain.
authors.affiliation: Department of Biochemistry and Molecular Biophysics, Washington University School of Medicine, St. Louis, Missouri 63110, USA.
authors.forename: Kanchan
authors.initials: K
authors.lastname: Garai
authors.forename: Sourajit M
authors.initials: SM
authors.lastname: Mustafi
authors.forename: Berevan
authors.initials: B
authors.lastname: Baban
authors.forename: Carl
authors.initials: C
authors.lastname: Frieden
mesh_terms: D053318:Apolipoprotein E3 / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D053327:Apolipoprotein E4 / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D005461:Fluorine / Q000737:chemistry; D008958:Models, Molecular; D009154:Mutation; D019906:Nuclear Magnetic Resonance, Biomolecular / Q000379:methods*; D011489:Protein Denaturation; D017434:Protein Structure, Tertiary; D014508:Urea / Q000737:chemistry
chemical_list: D053318:Apolipoprotein E3; D053327:Apolipoprotein E4; D005461:Fluorine; D014508:Urea