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Analysis by a highly sensitive split luciferase assay of the regions involved in APP dimerization and its impact on processing

Alzheimer’s disease (AD) is a neurodegenerative disease that causes progressive loss of cognitive functions, leading to dementia. Two types of lesions are found in AD brains: neurofibrillary tangles and senile plaques. The latter are composed mainly of the β-amyloid peptide (Aβ) generated by amyloid...

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Autores principales: Decock, Marie, El Haylani, Laetitia, Stanga, Serena, Dewachter, Ilse, Octave, Jean-Noël, Smith, Steven O., Constantinescu, Stefan N., Kienlen-Campard, Pascal
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2015
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4588712/
https://www.ncbi.nlm.nih.gov/pubmed/26500837
http://dx.doi.org/10.1016/j.fob.2015.09.002
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author Decock, Marie
El Haylani, Laetitia
Stanga, Serena
Dewachter, Ilse
Octave, Jean-Noël
Smith, Steven O.
Constantinescu, Stefan N.
Kienlen-Campard, Pascal
author_facet Decock, Marie
El Haylani, Laetitia
Stanga, Serena
Dewachter, Ilse
Octave, Jean-Noël
Smith, Steven O.
Constantinescu, Stefan N.
Kienlen-Campard, Pascal
author_sort Decock, Marie
collection PubMed
description Alzheimer’s disease (AD) is a neurodegenerative disease that causes progressive loss of cognitive functions, leading to dementia. Two types of lesions are found in AD brains: neurofibrillary tangles and senile plaques. The latter are composed mainly of the β-amyloid peptide (Aβ) generated by amyloidogenic processing of the amyloid precursor protein (APP). Several studies have suggested that dimerization of APP is closely linked to Aβ production. Nevertheless, the mechanisms controlling APP dimerization and their role in APP function are not known. Here we used a new luciferase complementation assay to analyze APP dimerization and unravel the involvement of its three major domains: the ectodomain, the transmembrane domain and the intracellular domain. Our results indicate that within cells full-length APP dimerizes more than its α and β C-terminal fragments, confirming the pivotal role of the ectodomain in this process. Dimerization of the APP transmembrane (TM) domain has been reported to regulate processing at the γ-cleavage site. We show that both non-familial and familial AD mutations in the TM GXXXG motifs strongly modulate Aβ production, but do not consistently change dimerization of the C-terminal fragments. Finally, we found for the first time that removal of intracellular domain strongly increases APP dimerization. Increased APP dimerization is linked to increased non-amyloidogenic processing.
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spelling pubmed-45887122015-10-23 Analysis by a highly sensitive split luciferase assay of the regions involved in APP dimerization and its impact on processing Decock, Marie El Haylani, Laetitia Stanga, Serena Dewachter, Ilse Octave, Jean-Noël Smith, Steven O. Constantinescu, Stefan N. Kienlen-Campard, Pascal FEBS Open Bio Research article Alzheimer’s disease (AD) is a neurodegenerative disease that causes progressive loss of cognitive functions, leading to dementia. Two types of lesions are found in AD brains: neurofibrillary tangles and senile plaques. The latter are composed mainly of the β-amyloid peptide (Aβ) generated by amyloidogenic processing of the amyloid precursor protein (APP). Several studies have suggested that dimerization of APP is closely linked to Aβ production. Nevertheless, the mechanisms controlling APP dimerization and their role in APP function are not known. Here we used a new luciferase complementation assay to analyze APP dimerization and unravel the involvement of its three major domains: the ectodomain, the transmembrane domain and the intracellular domain. Our results indicate that within cells full-length APP dimerizes more than its α and β C-terminal fragments, confirming the pivotal role of the ectodomain in this process. Dimerization of the APP transmembrane (TM) domain has been reported to regulate processing at the γ-cleavage site. We show that both non-familial and familial AD mutations in the TM GXXXG motifs strongly modulate Aβ production, but do not consistently change dimerization of the C-terminal fragments. Finally, we found for the first time that removal of intracellular domain strongly increases APP dimerization. Increased APP dimerization is linked to increased non-amyloidogenic processing. Elsevier 2015-09-06 /pmc/articles/PMC4588712/ /pubmed/26500837 http://dx.doi.org/10.1016/j.fob.2015.09.002 Text en © 2015 The Authors http://creativecommons.org/licenses/by-nc-nd/4.0/ This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).
spellingShingle Research article
Decock, Marie
El Haylani, Laetitia
Stanga, Serena
Dewachter, Ilse
Octave, Jean-Noël
Smith, Steven O.
Constantinescu, Stefan N.
Kienlen-Campard, Pascal
Analysis by a highly sensitive split luciferase assay of the regions involved in APP dimerization and its impact on processing
title Analysis by a highly sensitive split luciferase assay of the regions involved in APP dimerization and its impact on processing
title_full Analysis by a highly sensitive split luciferase assay of the regions involved in APP dimerization and its impact on processing
title_fullStr Analysis by a highly sensitive split luciferase assay of the regions involved in APP dimerization and its impact on processing
title_full_unstemmed Analysis by a highly sensitive split luciferase assay of the regions involved in APP dimerization and its impact on processing
title_short Analysis by a highly sensitive split luciferase assay of the regions involved in APP dimerization and its impact on processing
title_sort analysis by a highly sensitive split luciferase assay of the regions involved in app dimerization and its impact on processing
topic Research article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4588712/
https://www.ncbi.nlm.nih.gov/pubmed/26500837
http://dx.doi.org/10.1016/j.fob.2015.09.002
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