Abstract
gamma-Secretase is a membrane protein complex with an unusual aspartyl protease activity that catalyses the regulated intramembranous cleavage of the beta-amyloid precursor protein (APP) to release the Alzheimer's disease (AD)-associated amyloid beta-peptide (Abeta) and the APP intracellular domain (AICD). Here we show the reconstitution of gamma-secretase activity in the yeast Saccharomyces cerevisiae, which lacks endogenous gamma-secretase activity. Reconstituted gamma-secretase activity depends on the presence of four complex components including presenilin (PS), nicastrin (Nct), APH-1 (refs 3-6) and PEN-2 (refs 4, 7), is associated with endoproteolysis of PS, and produces Abeta and AICD in vitro. Thus, the biological activity of gamma-secretase is reconstituted by the co-expression of human PS, Nct, APH-1 and PEN-2 in yeast.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Alzheimer Disease / genetics
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Alzheimer Disease / metabolism
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Amyloid Precursor Protein Secretases
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Amyloid beta-Peptides / biosynthesis
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Amyloid beta-Protein Precursor / metabolism
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Aspartic Acid Endopeptidases
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Cell Membrane / enzymology*
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Cells, Cultured
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Endopeptidases / metabolism*
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Eukaryotic Cells / enzymology*
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Gene Expression Regulation, Enzymologic / genetics
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Gene Expression Regulation, Fungal / genetics
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Genes, Reporter / genetics
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Humans
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Macromolecular Substances
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Membrane Glycoproteins / metabolism
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Membrane Proteins / metabolism
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Peptide Hydrolases
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Presenilin-1
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Recombinant Fusion Proteins
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Saccharomyces cerevisiae / enzymology*
Substances
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Amyloid beta-Peptides
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Amyloid beta-Protein Precursor
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Macromolecular Substances
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Membrane Glycoproteins
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Membrane Proteins
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PSEN1 protein, human
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PSENEN protein, human
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Presenilin-1
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Recombinant Fusion Proteins
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nicastrin protein
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APH1A protein, human
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Amyloid Precursor Protein Secretases
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Endopeptidases
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Peptide Hydrolases
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Aspartic Acid Endopeptidases
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BACE1 protein, human