title: Biophysical properties of apolipoprotein E4 variants: implications in molecular mechanisms of correction of hypertriglyceridemia.
abstract: In humans and animal models, high plasma concentrations of apolipoprotein (apo) E are associated with hypertriglyceridemia. It has been shown that overexpression of human wild-type (WT) apoE4 in apoE-deficient mice induces hypertriglyceridemia. In contrast, overexpression of an apoE4 variant, apoE4-mut1 (apoE4(L261A, W264A, F265A, L268A, V269A)), does not induce hypertriglyceridemia and corrects hypercholesterolemia. Furthermore, overexpression of another variant, apoE4-mut2 (apoE4(W276A, L279A, V280A, V283A)), induces mild hypertriglyceridemia and does not correct hypercholesterolemia. To better understand how these mutations improve the function of apoE4, we investigated the conformation and stability of apoE4-mut1 and apoE4-mut2 and their binding to dimyristoyl phosphatidylcholine (DMPC) vesicles and to triglyceride (TG)-rich emulsion particles. We found that the mutations introduced in apoE4-mut1 lead to a more stable and compactly folded conformation of apoE4. These structural changes are associated with a slower rate of solubilization of DMPC vesicles by apoE4-mut1 and reduced binding of the protein to emulsion particles compared with WT apoE4. Under conditions of apoE4 overexpression, the reduced binding of apoE4-mut1 to TG-rich lipoprotein particles may facilitate the lipolysis of these particles and may alter the conformation of the lipoprotein-bound apoE in a way that favors the efficient clearance of the lipoprotein remnants. Mutations introduced in apoE4-mut2 result in smaller structural alterations compared with those observed in apoE4-mut1. The slightly altered structural properties of apoE4-mut2 are associated with slightly reduced binding of this protein to TG-rich lipoprotein particles and milder hypertriglyceridemia as compared with WT apoE4.
authors.affiliation: Department of Physiology and Biophysics, Boston University School of Medicine, 700 Albany Street, Boston, Massachusetts 02118, USA. igorshko@bu.edu
authors.forename: Irina N
authors.initials: IN
authors.lastname: Gorshkova
authors.forename: Kyriakos E
authors.initials: KE
authors.lastname: Kypreos
authors.forename: Donald L
authors.initials: DL
authors.lastname: Gantz
authors.forename: Vassilis I
authors.initials: VI
authors.lastname: Zannis
authors.forename: David
authors.initials: D
authors.lastname: Atkinson
mesh_terms: D000595:Amino Acid Sequence; D000817:Anilino Naphthalenesulfonates / Q000378:metabolism; D053327:Apolipoprotein E4 / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D002460:Cell Line; D002942:Circular Dichroism; D004134:Dimyristoylphosphatidylcholine / Q000737:chemistry / Q000378:metabolism; D006801:Humans; D015228:Hypertriglyceridemia / Q000378:metabolism*; D008079:Lipoproteins, VLDL / Q000378:metabolism; D008854:Microscopy, Electron; D008969:Molecular Sequence Data; D050505:Mutant Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D009154:Mutation; D011489:Protein Denaturation; D017510:Protein Folding; D055550:Protein Stability; D017433:Protein Structure, Secondary; D012995:Solubility; D013696:Temperature; D014304:Triolein / Q000378:metabolism
chemical_list: D000817:Anilino Naphthalenesulfonates; D053327:Apolipoprotein E4; D008079:Lipoproteins, VLDL; D050505:Mutant Proteins; D014304:Triolein; D004134:Dimyristoylphosphatidylcholine