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Metabolites of the β‐amyloid precursor protein generated by β‐secretase localise to the trans‐golgi network and late endosome in 293 cells
Author(s) -
Stephens David J.,
Austen Brian M.
Publication year - 1996
Publication title -
journal of neuroscience research
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.72
H-Index - 160
eISSN - 1097-4547
pISSN - 0360-4012
DOI - 10.1002/(sici)1097-4547(19961015)46:2<211::aid-jnr9>3.0.co;2-k
Subject(s) - endosome , golgi apparatus , amyloid precursor protein , amyloid precursor protein secretase , microbiology and biotechnology , chemistry , amyloid (mycology) , neuroscience , biology , alzheimer's disease , medicine , disease , endoplasmic reticulum , intracellular , inorganic chemistry
Deposition of β‐amyloid occurs in the brains of all sufferers of Alzheimer's disease. β‐amyloid is proteolytically derived from the β‐amyloid precursor protein by as yet unidentified enzymes termed secretases. We have generated and characterised antisera to the carboxy‐terminal domain and β‐secretase cleavage site of the Alzheimer's amyloid precursor protein. The β‐secretase cleavage event occurs at the extreme N‐terminus of the β‐amyloid peptide. Our antiserum to the N‐terminus of the β‐amyloid peptide (NTβ4) specifically recognises β‐secretase cleaved species as opposed to intact βAPP. NTβ4 specifically immunoprecipitates a 13 kDa fragment of βAPP (p13) which is potentially amyloidogenic. We have used these anti‐sera in confocal laser scanning immunofluorescence microscopy to localise the intracellular location of potentially amyloidogenic βAPP processing fragments such as p13. Using a number of marker antisera of known intracellular location, we have defined the major location of βAPP fragments possessing the Asp‐1 N‐terminus of β‐amyloid as the trans ‐Golgi network or late endosome on the basis of colocalisation with a monoclonal antibody to the cation‐independent mannose‐6‐phosphate receptor. The colocalisation was further investigated using brefeldin A which demonstrated that the p13 fragment and mannose‐6‐phosphate receptor are trafficked by alternative pathways from the trans ‐Golgi network. © 1996 Wiley‐Liss, Inc.