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

被引:112
作者
Fraering, PC
LaVoie, MJ
Ye, WJ
Ostaszewski, BL
Kimberly, WT
Selkoe, DJ
Wolfe, MS
机构
[1] Harvard Univ, Sch Med, Inst Med 730, Ctr Neurol Dis, Boston, MA 02115 USA
[2] Brigham & Womens Hosp, Ctr Neurol Dis, Boston, MA 02115 USA
关键词
D O I
10.1021/bi035748j
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
gamma-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 beta-protein precursor (APP) and the Notch receptor. Biochemical and genetic studies have identified four membrane proteins as components of gamma-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 gamma-secretase. Here we show that (1) the detergent dodecyl beta-D-maltoside (DDM) mediates the dissociation and inactivation of active gamma-secretase in a concentration-dependent manner, (2) DDM-dependent dissociation of the active gamma-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 gamma-secretase and offer a model for how the components of active gamma-secretase interact physically with each other.
引用
收藏
页码:323 / 333
页数:11
相关论文
共 48 条
[1]   The levels of, mature glycosylated nicastrin are regulated and correlate with γ-secretase processing of amyloid β-precursor protein [J].
Arawaka, S ;
Hasegawa, H ;
Tandon, A ;
Janus, C ;
Chen, FS ;
Yu, G ;
Kikuchi, K ;
Koyama, S ;
Kato, T ;
Fraser, PE ;
St George-Hyslop, P .
JOURNAL OF NEUROCHEMISTRY, 2002, 83 (05) :1065-1071
[2]   The proteolytic fragments of the Alzheimer's disease-associated presenilin-1 form heterodimers and occur as a 100-150-kDa molecular mass complex [J].
Capell, A ;
Grünberg, J ;
Pesold, B ;
Diehlmann, A ;
Citron, M ;
Nixon, R ;
Beyreuther, K ;
Selkoe, DJ ;
Haass, C .
JOURNAL OF BIOLOGICAL CHEMISTRY, 1998, 273 (06) :3205-3211
[3]   A presenilin-1-dependent γ-secretase-like protease mediates release of Notch intracellular domain [J].
De Strooper, B ;
Annaert, W ;
Cupers, P ;
Saftig, P ;
Craessaerts, K ;
Mumm, JS ;
Schroeter, EH ;
Schrijvers, V ;
Wolfe, MS ;
Ray, WJ ;
Goate, A ;
Kopan, R .
NATURE, 1999, 398 (6727) :518-522
[4]   Deficiency of presenilin-1 inhibits the normal cleavage of amyloid precursor protein [J].
De Strooper, B ;
Saftig, P ;
Craessaerts, K ;
Vanderstichele, H ;
Guhde, G ;
Annaert, W ;
Von Figura, K ;
Van Leuven, F .
NATURE, 1998, 391 (6665) :387-390
[5]   Presenilin and nicastrin regulate each other and determine amyloid β-peptide production via complex formation [J].
Edbauer, D ;
Winkler, E ;
Haass, C ;
Steiner, H .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 2002, 99 (13) :8666-8671
[6]   Reconstitution of γ-secretase activity [J].
Edbauer, D ;
Winkler, E ;
Regula, JT ;
Pesold, B ;
Steiner, H ;
Haass, C .
NATURE CELL BIOLOGY, 2003, 5 (05) :486-488
[7]   Activity-dependent isolation of the presenilin-γ-secretase complex reveals nicastrin and a γ substrate [J].
Esler, WP ;
Kimberly, WT ;
Ostaszewski, BL ;
Ye, WJ ;
Diehl, TS ;
Selkoe, DJ ;
Wolfe, MS .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 2002, 99 (05) :2720-2725
[8]   Transition-state analogue inhibitors of γ-secretase bind directly to presenilin-1 [J].
Esler, WP ;
Kimberly, WT ;
Ostaszewski, BL ;
Diehl, TS ;
Moore, CL ;
Tsai, JY ;
Rahmati, T ;
Xia, WM ;
Selkoe, DJ ;
Wolfe, MS .
NATURE CELL BIOLOGY, 2000, 2 (07) :428-434
[9]   Partial purification and characterization of γ-secretase from post-mortem human brain [J].
Farmery, MR ;
Tjernberg, LO ;
Pursglove, SE ;
Bergman, A ;
Winblad, B ;
Näslund, J .
JOURNAL OF BIOLOGICAL CHEMISTRY, 2003, 278 (27) :24277-24284
[10]   aph-1 and pen-2 are required for notch pathway signaling, γ-secretase cleavage of βAPP, and presenilin protein accumulation [J].
Francis, R ;
McGrath, G ;
Zhang, JH ;
Ruddy, DA ;
Sym, M ;
Apfeld, J ;
Nicoll, M ;
Maxwell, M ;
Hai, B ;
Ellis, MC ;
Parks, AL ;
Xu, W ;
Li, JH ;
Gurney, M ;
Myers, RL ;
Himes, CS ;
Hiebsch, R ;
Ruble, C ;
Nye, JS ;
Curtis, D .
DEVELOPMENTAL CELL, 2002, 3 (01) :85-97