Biogenesis and processing of the amyloid precursor protein in the early secretory pathway.
The beta-amyloid peptide is an aggregation-prone peptide that is released from the amyloid precursor protein (APP) after cleavage by the beta- and gamma-secretase. A number of studies have suggested that processing of APP by beta- and gamma-secretase occurs not only at the cell surface and in the endosomal compartments but also in the endoplasmic reticulum (ER) and Golgi complex. Here, we studied the role of the early secretory pathway in processing of APP. For this purpose, APP was in vitro translated in semi-permeabilized cells, which have a functionally intact ER and Golgi complex but lack a functional plasma membrane. We show that the beta-secretase cleavage product C99 is generated in the early secretory pathway. Moreover, nicastrin and presenilin, two components of the gamma-secretase complex, interacted with newly synthesized APP. Administration of the gamma-secretase inhibitor L685,458 caused accumulation of full length APP and C99. Full length APP but not C99 interacted with several protein quality control ER chaperones including the thiol oxidoreductase ERp57. Our in vitro study suggests that newly synthesized APP is subject to amyloidogenic processing during the initial phases of the secretory pathway.
Selivanova
Alexandra
A
Department of Neurobiology, Caring Sciences and Society (NVS), KI Alzheimer Disease Research Center, Karolinska Institutet, Novum 5th Floor, S-141 57 Stockholm, Sweden. alexandra.selivanova@ki.se
Winblad
Bengt
B
Dantuma
Nico P
NP
Farmery
Mark R
MR
D053829:Amyloid Precursor Protein Secretases / Q000378:metabolism*; D016564:Amyloid beta-Protein Precursor / Q000378:metabolism*; D002460:Cell Line; D004721:Endoplasmic Reticulum / Q000378:metabolism*; D006801:Humans; D007668:Kidney / Q000378:metabolism*; D014176:Protein Biosynthesis / Q000502:physiology; D015398:Signal Transduction / Q000502:physiology*
D016564:Amyloid beta-Protein Precursor; D053829:Amyloid Precursor Protein Secretases