title: Assembly of the gamma-secretase complex involves early formation of an intermediate subcomplex of Aph-1 and nicastrin.
abstract: The gamma-secretase complex is an unusual multimeric protease responsible for the intramembrane cleavage of a variety of type 1 transmembrane proteins, including the beta-amyloid precursor protein and Notch. Genetic and biochemical data have revealed that this protease consists of the presenilin heterodimer, a highly glycosylated form of nicastrin, and the recently identified gene products, Aph-1 and Pen-2. Whereas current evidence supports the notion that presenilin comprises the active site of the protease and that the other three components are members of the active complex required for proteolytic activity, the individual roles of the three co-factors remain unclear. Here, we demonstrate that endogenous Aph-1 interacts with an immature species of nicastrin, forming a stable intermediate early in the assembly of the gamma-secretase complex, prior to the addition of presenilin and Pen-2. Our data suggest 1) that Aph-1 is involved in the early stages of gamma-secretase assembly through the stabilization and perhaps glycosylation of nicastrin and by scaffolding nicastrin to the immature gamma-secretase complex, and 2) that presenilin, and later Pen-2, bind to this intermediate during the formation of the mature protease.
authors.affiliation: Center for Neurologic Diseases, Harvard Medical School and Brigham and Women's Hospital, Boston, Massachusetts 02115, USA.
authors.forename: Matthew J
authors.initials: MJ
authors.lastname: LaVoie
authors.forename: Patrick C
authors.initials: PC
authors.lastname: Fraering
authors.forename: Beth L
authors.initials: BL
authors.lastname: Ostaszewski
authors.forename: Wenjuan
authors.initials: W
authors.lastname: Ye
authors.forename: W Taylor
authors.initials: WT
authors.lastname: Kimberly
authors.forename: Michael S
authors.initials: MS
authors.lastname: Wolfe
authors.forename: Dennis J
authors.initials: DJ
authors.lastname: Selkoe
mesh_terms: D053829:Amyloid Precursor Protein Secretases; D000818:Animals; D016282:Aspartic Acid Endopeptidases; D016466:CHO Cells; D002793:Cholic Acids / Q000494:pharmacology; D006224:Cricetinae; D010450:Endopeptidases / Q000737:chemistry* / Q000502:physiology; D006801:Humans; D008562:Membrane Glycoproteins / Q000737:chemistry*; D008565:Membrane Proteins / Q000737:chemistry*; D010447:Peptide Hydrolases; D053764:Presenilin-1
chemical_list: D002793:Cholic Acids; D008562:Membrane Glycoproteins; D008565:Membrane Proteins; C508853:PSEN1 protein, human; C469349:PSENEN protein, human; D053764:Presenilin-1; C413604:nicastrin protein; C048531:chapso; C475154:APH1A protein, human; D053829:Amyloid Precursor Protein Secretases; D010450:Endopeptidases; D010447:Peptide Hydrolases; D016282:Aspartic Acid Endopeptidases; C509280:BACE1 protein, human