Detergent-Dependent Dissociation of Active γ-Secretase Reveals an Interaction between Pen-2 and PS1-NTF and Offers a Model for Subunit Organization within the Complex

Abstract

γ-Secretase is a member of a new class of proteases with an intramembrane catalytic site and cleaves numerous type I membrane proteins, including the amyloid β-protein precursor (APP) and the Notch receptor. Biochemical and genetic studies have identified four membrane proteins as components of γ-secretase: a heterodimeric form of presenilin (PS), composed of its N- and C-terminal fragments (PS-NTF and PS-CTF, respectively), a highly glycosylated, mature form of nicastrin (NCT), Aph-1, and Pen-2. However, it is unclear how these components interact physically with each other and assemble into functional complexes. We and others recently found that Aph-1 interacts with a less glycosylated, immature form of nicastrin as an intermediate toward full assembly of γ-secretase. Here we show that (1) the detergent dodecyl β-d-maltoside (DDM) mediates the dissociation and inactivation of active γ-secretase in a concentration-dependent manner, (2) DDM-dependent dissociation of the active γ-secretase complex generates two major inactive complexes (Pen-2−PS1-NTF and mNCT−Aph-1) and two minor inactive complexes (mNCT−Aph1−PS1-CTF and PS1-NTF−PS1-CTF), and (3) Pen-2 can also associate with the PS holoprotein in complexes devoid of NCT and Aph-1. Taken together, our results demonstrate that Pen-2 interacts with PS-NTF within active γ-secretase and offer a model for how the components of active γ-secretase interact physically with each other.