Amidation of beta-amyloid peptide strongly reduced the amyloidogenic activity without alteration of the neurotoxicity

J Neurochem. 1997 Nov;69(5):2048-54. doi: 10.1046/j.1471-4159.1997.69052048.x.

Abstract

Beta-amyloid accumulates in cerebral deposits in Alzheimer's disease, so to test the correlation between the neurotoxic and fibrillogenic capacity of beta-amyloid, we synthesized a peptide homologous to fragment 25-35 of beta-amyloid (beta25-35) and amidated at the C-terminus (beta25-35-NH2). As the amidation strongly reduced the amyloidogenic capacity of beta25-35, we compared its neurotoxic activity in the amidated (beta25-35-NH2) and nonamidated forms. The viability of primary cultures from fetal rat hippocampus was reduced in a dose-related manner (10-100 microM) similarly by beta25-35 and beta25-35-NH2, whereas a scrambled peptide, amidated or nonamidated, did not alter the neuronal viability. The neurotoxic activity of beta25-35-NH2 is mediated by apoptosis as demonstrated by morphological and biochemical investigations. Electron microscopy examination of culture media with beta25-35 or beta25-35-NH2 incubated with neuronal cells for 7 days confirmed the high level of fibrillogenic activity of beta25-35 and the almost total absence of fibrils in the solution with beta25-35-NH2. Furthermore, staining with thioflavine S was used to identify amyloid fibrils, and only the cultures exposed to beta25-35 exhibited intense staining associated with neuronal membranes. These data indicate that the neurotoxic activity of the beta-amyloid fragment is independent of the aggregated state of the peptide.

MeSH terms

  • Amino Acid Sequence
  • Amyloid beta-Peptides / chemistry
  • Amyloid beta-Peptides / toxicity*
  • Animals
  • Apoptosis / drug effects
  • Cell Nucleus / drug effects
  • Cell Nucleus / pathology
  • Cell Nucleus / ultrastructure
  • Cell Survival / drug effects
  • Cells, Cultured
  • Culture Media
  • DNA Fragmentation
  • Fetus
  • Hippocampus / cytology
  • Hippocampus / pathology*
  • Microscopy, Electron
  • Neurofibrils / drug effects
  • Neurofibrils / pathology
  • Neurofibrils / ultrastructure
  • Neurons / drug effects*
  • Neurons / pathology
  • Neurotoxins*
  • Peptide Fragments / chemistry
  • Peptide Fragments / toxicity*
  • Peptides / chemical synthesis
  • Peptides / chemistry
  • Peptides / toxicity
  • Rats

Substances

  • Amyloid beta-Peptides
  • Culture Media
  • Neurotoxins
  • Peptide Fragments
  • Peptides
  • amyloid beta-protein (25-35)