title: Self-association and stability of the ApoE isoforms at low pH: implications for ApoE-lipid interactions.
abstract: Apolipoprotein E (apoE) isoforms are known to differentially accumulate in the lysosomes of neuronal cells, and the deleterious effects of the apoE4 isoform in Alzheimer's disease may relate to its properties at the low lysosomal pH. However, the effect of pH on the molecular properties of full-length apoE is unclear. Here we examine the pH dependence of the monomer-dimer-tetramer reaction, of lipid binding, and of the stability of the three major apoE isoforms. Using FRET measurements, we find that the association-dissociation behavior of apoE proteins changes dramatically with changes in pH. At pH 4.5, approximating the pH of the lysosome, rate constants for association and dissociation are 2-10 times faster than those at pH 7.4. Aggregation beyond the tetrameric form is also more evident at lower pH values. Stability, as measured by urea denaturation at pH 4.5, is found to be considerably greater than that at neutral pH and to be isoform dependent. Lipid binding, as measured by turbidity clearance of unilamellar vesicles of DMPC, is faster at acidic pH values and consistent with our previous hypothesis that it is only the monomeric form of apoE that binds lipid tightly. Since apoE is more stable at pH 4.5 than at neutral pH, the more rapid apoE-lipid interactions at low pH are not correlated with the stability of the apoE isoforms, but rather to the faster association-dissociation behavior. Our results indicate that pathological behavior of apoE4 may arise from altered molecular properties of this protein at the acidic pH of the lysosome.
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: Berevan
authors.initials: B
authors.lastname: Baban
authors.forename: Carl
authors.initials: C
authors.lastname: Frieden
mesh_terms: D001057:Apolipoproteins E / Q000378:metabolism*; D006863:Hydrogen-Ion Concentration / Q000187:drug effects; D007700:Kinetics; D050356:Lipid Metabolism / Q000187:drug effects; D008958:Models, Molecular; D011489:Protein Denaturation / Q000187:drug effects; D020033:Protein Isoforms / Q000378:metabolism; D055550:Protein Stability / Q000187:drug effects; D020836:Protein Structure, Quaternary; D012965:Sodium Chloride / Q000494:pharmacology; D014508:Urea / Q000494:pharmacology
chemical_list: D001057:Apolipoproteins E; D020033:Protein Isoforms; D012965:Sodium Chloride; D014508:Urea