Site-specific modification of Alzheimer's peptides by cholesterol oxidation products enhances aggregation energetics and neurotoxicity.
Accumulation of amyloid beta-peptide (Abeta) and tau aggregates, possibly linked to age-associated deficiencies in protein homeostasis, appear to cause Alzheimer's disease. Schiff-base formation between Abeta and the aldehyde-bearing cholesterol oxidation product 3-beta-hydroxy-5-oxo-5,6-secocholestan-6-al is known to increase Abeta amyloidogenicity. Here, we synthesized Abeta variants site-specifically modified with the cholesterol aldehyde at Asp-1, Lys-16, or Lys-28, rather than studying mixtures. These distinct modifications have a similar effect on the thermodynamic propensity for aggregation, enabling aggregation at low concentrations. In contrast, the modification site differentially influences the aggregation kinetics; Lys-16-modified Abeta formed amorphous aggregates fastest and at the lowest concentration (within 2 h at a concentration of 20 nM), followed by the Lys-28 and Asp-1 conjugates. Also, the aggregates resulting from Abeta Lys-16 cholesterol aldehyde conjugation were more toxic to primary rat cortical neurons than treatment with unmodified Abeta under identical conditions and at the same concentration. Our results show that Abeta modification by cholesterol derivatives, especially at Lys-16, renders it kinetically and thermodynamically competent to form neurotoxic aggregates at concentrations approaching the physiologic concentration of Abeta.
Departments of Chemistry and Molecular and Experimental Medicine, Scripps Research Institute and Skaggs Institute for Chemical Biology, La Jolla, CA 92037, USA.
Kenji
K
Usui
John D
JD
Hulleman
Johan F
JF
Paulsson
Sarah J
SJ
Siegel
Evan T
ET
Powers
Jeffery W
JW
Kelly
D000544:Alzheimer Disease / Q000378:metabolism*; D000595:Amino Acid Sequence; D016229:Amyloid beta-Peptides / Q000737:chemistry* / Q000633:toxicity / Q000648:ultrastructure; D000818:Animals; D052160:Benzothiazoles; D002478:Cells, Cultured; D002784:Cholesterol / Q000378:metabolism*; D005453:Fluorescence; D007700:Kinetics; D008027:Light; D016253:Microscopy, Immunoelectron; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D009474:Neurons / Q000187:drug effects / Q000473:pathology*; D010084:Oxidation-Reduction / Q000187:drug effects; D010455:Peptides / Q000737:chemistry* / Q000633:toxicity*; D020836:Protein Structure, Quaternary; D051381:Rats; D012542:Scattering, Radiation; D013844:Thiazoles / Q000378:metabolism
D016229:Amyloid beta-Peptides; D052160:Benzothiazoles; D010455:Peptides; D013844:Thiazoles; C009462:thioflavin T; D002784:Cholesterol