Abstract
Presenilin (PS) mutations are associated with early-onset Alzheimer's disease and PS proteins are involved with gamma-secretase cleavage of the amyloid precursor protein, APP. We have shown previously that alpha-, beta- and gamma-secretase cleavages of APP are conserved in Pichia pastoris. Here, we report co-expression of APP and PSI in P. pastoris and show by immunoelectron microscopy colocalization of these two proteins in expanded endoplasmic reticulum. Western blot analysis indicates a drastic reduction of both alpha- and beta-secretase products. A relative increase in beta-secretase product derived from immature APP is also observed, pointing to a beta-secretase activity of P. pastoris associated with the early secretory pathway.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Amyloid Precursor Protein Secretases
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Amyloid beta-Protein Precursor / biosynthesis
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Amyloid beta-Protein Precursor / genetics
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Amyloid beta-Protein Precursor / metabolism*
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Aspartic Acid Endopeptidases / metabolism
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Blotting, Western
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Clone Cells / cytology
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Clone Cells / enzymology
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Clone Cells / metabolism
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Endopeptidases
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Endoplasmic Reticulum / metabolism
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Glycosylation
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Immunohistochemistry
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Membrane Proteins / biosynthesis*
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Membrane Proteins / genetics*
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Microscopy, Electron
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Models, Biological
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Pichia / cytology
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Pichia / enzymology*
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Pichia / genetics*
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Pichia / ultrastructure
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Presenilin-1
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Protein Processing, Post-Translational
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Transfection
Substances
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Amyloid beta-Protein Precursor
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Membrane Proteins
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Presenilin-1
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Amyloid Precursor Protein Secretases
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Endopeptidases
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Aspartic Acid Endopeptidases