Formation of amyloid-like fibrils in COS cells overexpressing part of the Alzheimer amyloid protein precursor

Nature. 1990 Oct 11;347(6293):566-9. doi: 10.1038/347566a0.

Abstract

A pathological hallmark of Alzheimer's disease is the deposition of amyloid fibrils in the brain. The principal component of the amyloid fibril is beta/A4 protein, which is derived from a large membrane-bound glycoprotein, Alzheimer amyloid protein precursor (APP). Although the deposition of amyloid is thought to result from the aberrant processing of APP, the detailed molecular mechanisms of amyloidogenesis remain unclear. A C-terminal fragment of APP which spans the beta/A4 and cytoplasmic domains has a tendency to self-aggregate. In an attempt to establish a cultured-cell model for amyloid fibril formation, we have transfected COS-1 cells with complementary DNA encoding the C-terminal 100 residues of APP. In the perinuclear regions of a small population of DNA-transfected cells, we observed inclusion-like deposits which showed a strong immunohistochemical reaction towards an anti-C-terminal APP antibody or an anti-beta/A4 amyloid core-specific antibody. Electron microscope observations of the inclusion-carrying cells revealed an accumulation of amyloid-like fibrils of 8-22 nm diameter near and on the nuclear membrane. The fibrils showed a beaded or helical structure, and reacted positively with the anti-C-terminus antibody by immunoelectron microscopy. These results suggest that the formation of amyloid fibrils is an inherent characteristic of the C-terminal peptide of APP. The present system provides a suitable model for the molecular dissection of the process of brain amyloidogenesis.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Alzheimer Disease / metabolism*
  • Amyloid beta-Peptides / genetics*
  • Amyloid beta-Peptides / metabolism*
  • Amyloid beta-Protein Precursor
  • Animals
  • Base Sequence
  • Cell Line
  • Chlorocebus aethiops
  • Cloning, Molecular
  • DNA / genetics
  • Gene Expression*
  • Immunoenzyme Techniques
  • Immunohistochemistry
  • Inclusion Bodies / metabolism
  • Macromolecular Substances
  • Microscopy, Electron
  • Microscopy, Immunoelectron
  • Molecular Sequence Data
  • Peptide Fragments / genetics
  • Protein Precursors / genetics*
  • Transfection

Substances

  • Amyloid beta-Peptides
  • Amyloid beta-Protein Precursor
  • Macromolecular Substances
  • Peptide Fragments
  • Protein Precursors
  • DNA