title: Specific Binding of Cholesterol to C99 Domain of Amyloid Precursor Protein Depends Critically on Charge State of Protein.
abstract: Recent NMR chemical shift measurements of the 99 residue C-terminal fragment of amyloid precursor protein (APP-C99) in the presence of cholesterol provide evidence of binary complex formation between C99 and cholesterol in membrane mimetic environments. It has also been observed that the production of Aβ protein is enhanced under conditions of high cholesterol concentration. In this study, we investigated the impact of the charge state of C99 on the structure and stability of the C99-cholesterol complex. We observed that the binding of C99 to cholesterol depends critically on the charge state of Glu 693 (E22) and Asp 694 (D23). Evaluation of the pKa values of the Asp and Glu side chains suggests that these residues may be predominantly neutral in existing experimental observations of a stable C99-cholesterol complex at lower pH (characteristic of the endosomal environment), while binding is destabilized near neutral pH (characteristic of the cytoplasm). These observations suggest that specific binding of cholesterol to C99 is a sensitive function of the pH encountered in vivo, with key E22 and D23 residues serving as a "pH switch" controlling C99-cholesterol binding.
authors.affiliation: Department of Chemistry, Boston University , 590 Commonwealth Avenue, Boston, Massachusetts 02215, United States.
authors.forename: Afra
authors.initials: A
authors.lastname: Panahi
authors.affiliation: Department of Chemistry, Boston University , 590 Commonwealth Avenue, Boston, Massachusetts 02215, United States.
authors.forename: Asanga
authors.initials: A
authors.lastname: Bandara
authors.affiliation: Department of Chemistry, Boston University , 590 Commonwealth Avenue, Boston, Massachusetts 02215, United States.
authors.forename: George A
authors.initials: GA
authors.lastname: Pantelopulos
authors.affiliation: Biophysical Chemistry Laboratory, Physical Chemistry Department, School of Chemistry, National Autonomous University of Mexico , Mexico City, Mexico.
authors.forename: Laura
authors.initials: L
authors.lastname: Dominguez
authors.affiliation: Department of Chemistry, Boston University , 590 Commonwealth Avenue, Boston, Massachusetts 02215, United States.
authors.forename: John E
authors.initials: JE
authors.lastname: Straub
mesh_terms: D016564:Amyloid beta-Protein Precursor / Q000737:chemistry*; D002784:Cholesterol / Q000737:chemistry*; D006801:Humans; D008051:Lipid Bilayers / Q000737:chemistry*; D011485:Protein Binding; D011506:Proteins / Q000737:chemistry*
chemical_list: D016564:Amyloid beta-Protein Precursor; D008051:Lipid Bilayers; D011506:Proteins; D002784:Cholesterol