Enhanced toxicity and cellular binding of a modified amyloid beta peptide with a methionine to valine substitution.
The amyloid beta peptide (Abeta) is toxic to neuronal cells, and it is probable that this toxicity is responsible for the progressive cognitive decline associated with Alzheimer's disease. However, the nature of the toxic Abeta species and its precise mechanism of action remain to be determined. It has been reported that the methionine residue at position 35 has a pivotal role to play in the toxicity of Abeta. We examined the effect of mutating the methionine to valine in Abeta42 (AbetaM35V). The neurotoxic activity of AbetaM35V on primary mouse neuronal cortical cells was enhanced, and this diminished cell viability occurred at an accelerated rate compared with Abeta42. AbetaM35V binds Cu2+ and produces similar amounts of H2O2 as Abeta42 in vitro, and the neurotoxic activity was attenuated by the H2O2 scavenger catalase. The increased toxicity of AbetaM35V was associated with increased binding of this mutated peptide to cortical cells. The M35V mutation altered the interaction between Abeta and copper in a lipid environment as shown by EPR analysis, which indicated that the valine substitution made the peptide less rigid in the bilayer region with a resulting higher affinity for the bilayer. Circular dichroism spectroscopy showed that both Abeta42 and AbetaM35V displayed a mixture of alpha-helical and beta-sheet conformations. These findings provide further evidence that the toxicity of Abeta is regulated by binding to neuronal cells.
Department of Pathology, University of Melbourne, Victoria 3010, Australia.
Giuseppe D
GD
Ciccotosto
Deborah
D
Tew
Cyril C
CC
Curtain
Danielle
D
Smith
Darryl
D
Carrington
Colin L
CL
Masters
Ashley I
AI
Bush
Robert A
RA
Cherny
Roberto
R
Cappai
Kevin J
KJ
Barnham
D016229:Amyloid beta-Peptides / Q000737:chemistry* / Q000235:genetics* / Q000378:metabolism; D000818:Animals; D002374:Catalase / Q000378:metabolism; D002384:Catalysis; D002470:Cell Survival; D002478:Cells, Cultured; D002540:Cerebral Cortex / Q000378:metabolism; D002942:Circular Dichroism; D003300:Copper / Q000737:chemistry; D004305:Dose-Response Relationship, Drug; D004578:Electron Spin Resonance Spectroscopy; D006861:Hydrogen Peroxide / Q000494:pharmacology; D007700:Kinetics; D008051:Lipid Bilayers / Q000737:chemistry; D050356:Lipid Metabolism; D008715:Methionine / Q000737:chemistry* / Q000235:genetics; D051379:Mice; D008854:Microscopy, Electron; D009154:Mutation; D009474:Neurons / Q000378:metabolism; D010455:Peptides / Q000737:chemistry; D011485:Protein Binding; D011487:Protein Conformation; D017433:Protein Structure, Secondary; D013997:Time Factors; D014633:Valine / Q000737:chemistry* / Q000235:genetics
D016229:Amyloid beta-Peptides; D008051:Lipid Bilayers; D010455:Peptides; D003300:Copper; D008715:Methionine; D006861:Hydrogen Peroxide; D002374:Catalase; D014633:Valine