Generation of Alzheimer β-amyloid protein in the trans-Golgi network in the apparent absence of vesicle formation

Huaxi Xu, David Sweeney, Rong Wang, Gopal Thinakaran, Amy C.Y. Lo, Sangram S. Sisodia, Paul Greengard, Sam Gandy

Research output: Contribution to journalArticlepeer-review

256 Scopus citations

Abstract

β-amyloid protein (Aβ) formation was reconstituted in permeabilized neuroblastoma cells expressing human Alzheimer β-amyloid precursor protein (βAPP) harboring the Swedish double mutation associated with familial early- onset Alzheimer disease. Permeabilized cells were prepared following metabolic labeling and incubation at 20°C, a temperature that allows βAPP to accumulate in the trans-Golgi network (TGN) without concomitant Aβ formation. Subsequent incubation at 37°C led to the generation of Aβ. Aβ production in the TGN persisted even under conditions in which formation of nascent post-TGN vesicles was inhibited by addition of guanosine 5'-O-(3- thiotriphosphate), a nonhydrolyzable GTP analogue, or by omission of cytosol. These and other results indicate that vesicle budding and trafficking may not be required for proteolytic metabolism of βAPP to Aβ, a process that includes 'γ-secretase' cleavage within the βAPP transmembrane domain.

Original languageEnglish
Pages (from-to)3748-3752
Number of pages5
JournalProceedings of the National Academy of Sciences of the United States of America
Volume94
Issue number8
DOIs
StatePublished - 15 Apr 1997
Externally publishedYes

Fingerprint

Dive into the research topics of 'Generation of Alzheimer β-amyloid protein in the trans-Golgi network in the apparent absence of vesicle formation'. Together they form a unique fingerprint.

Cite this