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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.
- Source :
-
Journal of neurochemistry [J Neurochem] 2001 Mar; Vol. 76 (5), pp. 1308-14. - Publication Year :
- 2001
-
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.
- Subjects :
- Amyloid beta-Protein Precursor biosynthesis
Animals
DNA, Complementary
Green Fluorescent Proteins
Humans
Luminescent Proteins analysis
Luminescent Proteins genetics
Molecular Weight
Neuroblastoma
Protein Isoforms biosynthesis
Protein Isoforms genetics
Rats
Recombinant Fusion Proteins biosynthesis
Subcellular Fractions metabolism
Subcellular Fractions ultrastructure
Transfection
Tumor Cells, Cultured
Alzheimer Disease genetics
Amyloid beta-Protein Precursor genetics
RNA, Messenger genetics
Sequence Deletion
Subjects
Details
- Language :
- English
- ISSN :
- 0022-3042
- Volume :
- 76
- Issue :
- 5
- Database :
- MEDLINE
- Journal :
- Journal of neurochemistry
- Publication Type :
- Academic Journal
- Accession number :
- 11238715
- Full Text :
- https://doi.org/10.1046/j.1471-4159.2001.00122.x