Alzheimer's Abeta peptides containing an isostructural backbone mutation afford distinct aggregate morphologies but analogous cytotoxicity. Evidence for a common low-abundance toxic structure(s)?
Amyloid beta (Abeta) peptide amyloidogenesis, involving the formation of numerous distinct quaternary structures, appears to cause Alzheimer's disease. However, the precise identification of the toxic structure(s) and the neurotoxicity mechanism(s) remains elusive. Mutating the Abeta 1-40 Phe19-Phe20 backbone amide bond to an isostructural E-olefin bond enables formation of spherical aggregates to the exclusion of detectable amyloid fibrils. Herein, the fibrillization and toxicity of amide-to-ester mutants of Abeta 1-40 at the 19-20 position and surrounding backbone amide bonds are compared to the fibrillization and toxicity of the 19-20 E-olefin Abeta analogue and wild type Abeta. Whereas isostructural amide-to-E-olefin mutations eliminate both the H-bond donor and acceptor capabilities, isostructural amide-to-ester mutations eliminate the donor while retaining the ester carbonyl as a weakened acceptor. None of the amide-to-ester Abeta 1-40 mutants prevent fibrillization; in fact several exhibit hastened amyloidogenesis. The 18-19 amide-to-ester substitution is the only backbone mutation within the hydrophobic core region of the fibril (residues 17-21) that significantly slows fibrillization. Despite forming different morphologies, the 19-20 E-olefin mutant, the 18-19 amide-to-ester mutant, and WT Abeta 1-40 fibrils all exhibit similar toxicities when applied to PC12 cells at 18 h into the aggregation reactions, as assessed by MTT metabolic activity measurements. This result suggests that a common but low abundance aggregate morphology, that is accessible to these Abeta analogues, mediates toxicity, or that several different aggregate morphologies are similarly toxic.
Bieschke
Jan
J
Department of Chemistry and The Skaggs Institute for Chemical Biology, The Scripps Research Institute, 10550 North Torrey Pines Road, La Jolla, California 92037, USA.
Siegel
Sarah J
SJ
Fu
Yanwen
Y
Kelly
Jeffery W
JW
D000544:Alzheimer Disease / Q000378:metabolism; D000577:Amides / Q000737:chemistry; D000682:Amyloid / Q000737:chemistry / Q000235:genetics; D016229:Amyloid beta-Peptides / Q000737:chemistry* / Q000235:genetics; D000818:Animals; D045744:Cell Line, Tumor; D002942:Circular Dichroism; D004952:Esters / Q000737:chemistry; D006860:Hydrogen Bonding; D018625:Microscopy, Atomic Force; D008855:Microscopy, Electron, Scanning; D008958:Models, Molecular; D018716:Molecular Mimicry; D050505:Mutant Proteins / Q000737:chemistry* / Q000235:genetics; D009154:Mutation; D010455:Peptides / Q000138:chemical synthesis / Q000737:chemistry* / Q000235:genetics; D051381:Rats
D000577:Amides; D000682:Amyloid; D016229:Amyloid beta-Peptides; D004952:Esters; D050505:Mutant Proteins; D010455:Peptides