Glu-333 of nicastrin directly participates in γ-secretase activity

Daniel R. Dries, Sanjiv Shah, Yu Hong Han, Cong Yu, Sophie Yu, Mark S. Shearman, Gang Yu

Research output: Contribution to journalArticle

49 Citations (Scopus)

Abstract

γ-Secretase is a proteolytic membrane complex that processes a variety of substrates including the amyloid precursor protein and the Notch receptor. Earlier we showed that one of the components of this complex, nicastrin (NCT), functions as a receptor for γ-secretase substrates. A recent report challenged this, arguing instead that the Glu-333 residue of NCT predicted to participate in substrate recognition only participates in γ-secretase complex maturation and not in activity per se. Here, we present evidence that Glu-333 directly participates in γ-secretase activity. By normalizing to the active pool of γ-secretase with two separate methods, we establish that γ-secretase complexes containing NCT-E333A are indeed deficient in intrinsic activity. We also demonstrate that the NCT-E333A mutant is deficient in its binding to substrates. Moreover, we find that the cleavage of substrates by γ-secretase activity requires a free N-terminal amine but no minimal length of the extracellular N-terminal stub. Taken together, these studies provide further evidence supporting the role of NCT in substrate recognition. Finally, because γ-secretase cleaves itself during its maturation and because NCT-E333A also shows defects in γ-secretase complex maturation, we present a model whereby Glu-333 can serve a dual role via similar mechanisms in the recruitment of both Type 1 membrane proteins for activity and the presenilin intracellular loop during complex maturation.

Original languageEnglish (US)
Pages (from-to)29714-29724
Number of pages11
JournalJournal of Biological Chemistry
Volume284
Issue number43
DOIs
StatePublished - Oct 23 2009

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Amyloid Precursor Protein Secretases
Substrates
Presenilins
Notch Receptors
Amyloid beta-Protein Precursor
Amines
Membrane Proteins
Membranes
Defects

ASJC Scopus subject areas

  • Biochemistry
  • Cell Biology
  • Molecular Biology

Cite this

Dries, D. R., Shah, S., Han, Y. H., Yu, C., Yu, S., Shearman, M. S., & Yu, G. (2009). Glu-333 of nicastrin directly participates in γ-secretase activity. Journal of Biological Chemistry, 284(43), 29714-29724. https://doi.org/10.1074/jbc.M109.038737

Glu-333 of nicastrin directly participates in γ-secretase activity. / Dries, Daniel R.; Shah, Sanjiv; Han, Yu Hong; Yu, Cong; Yu, Sophie; Shearman, Mark S.; Yu, Gang.

In: Journal of Biological Chemistry, Vol. 284, No. 43, 23.10.2009, p. 29714-29724.

Research output: Contribution to journalArticle

Dries, DR, Shah, S, Han, YH, Yu, C, Yu, S, Shearman, MS & Yu, G 2009, 'Glu-333 of nicastrin directly participates in γ-secretase activity', Journal of Biological Chemistry, vol. 284, no. 43, pp. 29714-29724. https://doi.org/10.1074/jbc.M109.038737
Dries DR, Shah S, Han YH, Yu C, Yu S, Shearman MS et al. Glu-333 of nicastrin directly participates in γ-secretase activity. Journal of Biological Chemistry. 2009 Oct 23;284(43):29714-29724. https://doi.org/10.1074/jbc.M109.038737
Dries, Daniel R. ; Shah, Sanjiv ; Han, Yu Hong ; Yu, Cong ; Yu, Sophie ; Shearman, Mark S. ; Yu, Gang. / Glu-333 of nicastrin directly participates in γ-secretase activity. In: Journal of Biological Chemistry. 2009 ; Vol. 284, No. 43. pp. 29714-29724.
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