Uncoupled packaging of amyloid precursor protein and presenilin 1 into coat protein complex II vesicles

Jinoh Kim, Susan Hamamoto, Mariella Ravazzola, Lelio Orci, Randy Schekman

Research output: Contribution to journalArticle

67 Citations (Scopus)

Abstract

Mutant forms of presenilin (PS) 1 and 2 and amyloid precursor protein (APP) lead to familial Alzheimer's disease. Several reports indicate that PS may modulate APP export from the endoplasmic reticulum (ER). To develop a test of this possibility, we reconstituted the capture of APP and PS1 in COPII (coat protein complex II) vesicles formed from ER membranes in permeabilized cultured cells. The recombinant forms of mammalian COPII proteins were active in a reaction that measures coat subunit assembly and coated vesicle budding on chemically defined synthetic liposomes. However, the recombinant COPII proteins were not active in cargo capture and vesicle budding from microsomal membranes. In contrast, rat liver cytosol was active in stimulating the sorting and packaging of APP, PS1, and p58 (an itinerant ER to Golgi marker protein) into transport vesicles from donor ER membranes. Budding was stimulated in dilute cytosol by the addition of recombinant COPII proteins. Fractionation of the cytosol suggested one or more additional proteins other than the COPII snbunits may be essential for cargo selection or vesicle formation from the mammalian ER membrane. The recombinant Sec24C specifically recognized the APP C-terminal region for packaging. Titration of Sarla distinguished the packaging requirements of APP and PS1. Furthermore, APP packaging was not affected by deletion of PS1 or PS1 and 2, suggesting APP and PS1 trafficking from the ER are normally uncoupled.

Original languageEnglish (US)
Pages (from-to)7758-7768
Number of pages11
JournalJournal of Biological Chemistry
Volume280
Issue number9
DOIs
StatePublished - Mar 4 2005

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Presenilin-1
Amyloid beta-Protein Precursor
Capsid Proteins
Product Packaging
Endoplasmic Reticulum
Packaging
Cytosol
Membranes
Recombinant proteins
Protein Transport
Recombinant Proteins
Amyloid
Proteins
Presenilin-2
Presenilins
Coated Vesicles
Transport Vesicles
Protein C
Liposomes
Fractionation

ASJC Scopus subject areas

  • Biochemistry

Cite this

Uncoupled packaging of amyloid precursor protein and presenilin 1 into coat protein complex II vesicles. / Kim, Jinoh; Hamamoto, Susan; Ravazzola, Mariella; Orci, Lelio; Schekman, Randy.

In: Journal of Biological Chemistry, Vol. 280, No. 9, 04.03.2005, p. 7758-7768.

Research output: Contribution to journalArticle

Kim, Jinoh ; Hamamoto, Susan ; Ravazzola, Mariella ; Orci, Lelio ; Schekman, Randy. / Uncoupled packaging of amyloid precursor protein and presenilin 1 into coat protein complex II vesicles. In: Journal of Biological Chemistry. 2005 ; Vol. 280, No. 9. pp. 7758-7768.
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