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Direct visualization of the gamma secretase‐generated carboxyl‐terminal domain of the amyloid precursor protein: association with Fe65 and translocation to the nucleus
Author(s) -
Kinoshita A.,
Whelan C. M.,
Smith C. J.,
Berezovska O.,
Hyman B. T.
Publication year - 2002
Publication title -
journal of neurochemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.75
H-Index - 229
eISSN - 1471-4159
pISSN - 0022-3042
DOI - 10.1046/j.1471-4159.2002.01016.x
Subject(s) - amyloid precursor protein , nucleus , cytoplasm , chemistry , alpha secretase , förster resonance energy transfer , amyloid precursor protein secretase , cleavage (geology) , green fluorescent protein , microbiology and biotechnology , p3 peptide , biophysics , colocalization , bimolecular fluorescence complementation , biochemistry , alzheimer's disease , fluorescence , biology , medicine , paleontology , physics , disease , pathology , quantum mechanics , fracture (geology) , gene , yeast
Amyloid‐β, the peptide that deposits as senile plaques in Alzheimer's disease, is derived from the amyloid precursor protein (APP) by a gamma secretase‐mediated intramembranous cleavage. In addition to amyloid‐β, this cleavage produces a carboxyl‐terminal intracellular fragment which has an unknown function. The carboxyl‐terminal domain of APP interacts in the cytoplasm with an adapter protein, Fe65. We demonstrate by laser scanning confocal microscopy that a gamma secretase generated APP carboxyl‐terminal domain, tagged with green fluorescent protein (GFP), translocates to the nucleus in a manner dependent upon stabilization by the adapter protein Fe65; APP which has been mutated to block interactions with Fe65 cannot be detected in the nucleus. The APP‐CT domain continues to interact with Fe65 in the nucleus, as determined by both colocalization and fluorescence resonance energy transfer (FRET). Visualization of the APP‐CT‐Fe65 complex in the nucleus may serve as a readout for processes that modify gamma secretase release of APP‐CT.