Novel β-secretase cleavage of β-amyloid precursor protein in the endoplasmic reticulum/intermediate compartment of NT2N cells

Abraham S.C. Chyung, Barry D. Greenberg, David G. Cook, Robert W. Doms, Virginia M.Y. Lee

Research output: Contribution to journalArticlepeer-review

Abstract

Previous studies have demonstrated that NT2N neurons derived from a human embryonal carcinoma cell line (NT2) constitutively process the endogenous wild-type β-amyloid precursor protein (APP) to amyloid β peptide in an intracellular compartment. These studies indicate that other proteolytic fragments generated by intracellular processing must also be present in these cells. Here we show that the NH2-terminal fragment of APP generated by β-secretase cleavage (APPβ) is indeed produced from the endogenous full length APP (APP(FL)). Pulse-chase studies demonstrated a precursor-product relationship between APP(FL) and APPβ as well as intracellular and secreted APPβ fragments. In addition, trypsin digestion of intact NT2N cells at 4°C did not abolish APPβ recovered from the cell lysates. Furthermore, the production of intracellular APPβ from wild-type APP appears to be a unique characteristic of postmitotic neurons, since intracellular APPβ was not detected in several non-neuronal cell lines. Significantly, production of APPβ occurred even when APP was retained in the ER/intermediate compartment by inhibition with brefeldin A, incubation at 15°C, or by expression of exogenous APP bearing the dilysine ER retrieval motif.

Original languageEnglish (US)
Pages (from-to)671-680
Number of pages10
JournalJournal of Cell Biology
Volume138
Issue number3
DOIs
StatePublished - Aug 11 1997
Externally publishedYes

ASJC Scopus subject areas

  • Cell Biology

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