title: Phosphate and HEPES buffers potently affect the fibrillation and oligomerization mechanism of Alzheimer's Aβ peptide.
abstract: The oligomerization of Aβ peptide into amyloid fibrils is a hallmark of Alzheimer's disease. Due to its biological relevance, phosphate is the most commonly used buffer system for studying the formation of Aβ and other amyloid fibrils. Investigation into the characteristics and formation of amyloid fibrils frequently relies upon material formed in vitro, predominantly in phosphate buffers. Herein, we examine the effects on the fibrillation and oligomerization mechanism of Aβ peptide that occur due solely to the influence of phosphate buffer. We reveal that significant differences in amyloid fibrillation are observed due to fibrillation being initiated in phosphate or HEPES buffer (at physiological pH and temperature). Except for the differing buffer ions, all experimental parameters were kept constant. Fibril formation was assessed using fluorescently monitored kinetic studies, microscopy, X-ray fiber diffraction and infrared and nuclear magnetic resonance spectroscopies. Based on this set up, we herein reveal profound effects on the mechanism and speed of Aβ fibrillation. The three histidine residues at positions 6, 13 and 14 of Aβ(1-40) are instrumental in these mechanistic changes. We conclude that buffer plays a more significant role in fibril formation than has been generally acknowledged.
authors.affiliation: Max-Planck-Forschungsstelle für Enzymologie der Proteinfaltung, Weinbergweg 22, D-06120 Halle (Saale), Germany.
authors.forename: Megan
authors.initials: M
authors.lastname: Garvey
authors.forename: Katharina
authors.initials: K
authors.lastname: Tepper
authors.forename: Caroline
authors.initials: C
authors.lastname: Haupt
authors.forename: Uwe
authors.initials: U
authors.lastname: Knüpfer
authors.forename: Karolin
authors.initials: K
authors.lastname: Klement
authors.forename: Jessica
authors.initials: J
authors.lastname: Meinhardt
authors.forename: Uwe
authors.initials: U
authors.lastname: Horn
authors.forename: Jochen
authors.initials: J
authors.lastname: Balbach
authors.forename: Marcus
authors.initials: M
authors.lastname: Fändrich
mesh_terms: D000544:Alzheimer Disease / Q000378:metabolism*; D000682:Amyloid / Q000737:chemistry*; D016229:Amyloid beta-Peptides / Q000737:chemistry*; D002021:Buffers; D006531:HEPES / Q000737:chemistry*; D006639:Histidine / Q000737:chemistry; D006801:Humans; D010446:Peptide Fragments / Q000737:chemistry*; D010710:Phosphates / Q000737:chemistry*
chemical_list: D000682:Amyloid; D016229:Amyloid beta-Peptides; D002021:Buffers; D010446:Peptide Fragments; D010710:Phosphates; C078592:amyloid beta-protein (1-40); D006639:Histidine; D006531:HEPES