A dinucleotide deletion in amyloid precursor protein (APP) mRNA associated with sporadic Alzheimer's disease results in efficient secretion of truncated APP isoforms from neuroblastoma cell cultures

J Neurochem. 2001 Mar;76(5):1308-14. doi: 10.1046/j.1471-4159.2001.00122.x.

Abstract

Recently, two dinucleotide deletions were detected in the mRNA of the amyloid precursor protein (APP) from cerebral cortex neurons of patients with sporadic Alzheimer's disease (AD) or Down's syndrome. These deletions resulted in truncation of APP, producing an APP isoform with a 38-kDa N-terminus and a novel carboxyl terminus (APP+1). We investigated the subcellular localization and the processing of APP+1 in the neuroblastoma cell line B103. cDNA constructs were generated encoding fusion proteins of APP+1 or full-length APP with the enhanced green fluorescent protein (eGFP). In transient transfection experiments using B103 cells, the APP+1-eGFP fusion protein showed a reticular localization with intense staining in the Golgi complex. Unlike full-length APP fused to eGFP, the APP+1-eGFP fusion protein did not localize to the perinuclear area or to the plasma membrane. Western blot analysis of cell extracts confirmed the translation of the expected fusion proteins. Analysis of the supernatant by western blot indicated that the APP+1-eGFP fusion protein was efficiently secreted from B103 cells, whereas the secreted form of full-length APP fusion protein (APPs) was hardly detectable. Thus, both dinucleotide deletions in the APP mRNA result in truncated APP+1 that is not membrane associated and is readily secreted from neurons.

Publication types

  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Alzheimer Disease / genetics*
  • Amyloid beta-Protein Precursor / biosynthesis
  • Amyloid beta-Protein Precursor / genetics*
  • Animals
  • DNA, Complementary
  • Green Fluorescent Proteins
  • Humans
  • Luminescent Proteins / analysis
  • Luminescent Proteins / genetics
  • Molecular Weight
  • Neuroblastoma
  • Protein Isoforms / biosynthesis
  • Protein Isoforms / genetics
  • RNA, Messenger / genetics*
  • Rats
  • Recombinant Fusion Proteins / biosynthesis
  • Sequence Deletion*
  • Subcellular Fractions / metabolism
  • Subcellular Fractions / ultrastructure
  • Transfection
  • Tumor Cells, Cultured

Substances

  • Amyloid beta-Protein Precursor
  • DNA, Complementary
  • Luminescent Proteins
  • Protein Isoforms
  • RNA, Messenger
  • Recombinant Fusion Proteins
  • Green Fluorescent Proteins