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P2‐363: Increased catalytic autoantibodies to amyloid‐β peptide in Alzheimer's disease
Author(s) -
Taguchi Hiroaki,
Planque Stephanie,
Nishiyama Yasuhiro,
Szabo Paul,
Weksler Mark E.,
Friedland Robert P.,
Paul Sudhir
Publication year - 2008
Publication title -
alzheimer's and dementia
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 6.713
H-Index - 118
eISSN - 1552-5279
pISSN - 1552-5260
DOI - 10.1016/j.jalz.2008.05.1441
Subject(s) - antigen , chemistry , autoantibody , peptide , serine protease , antibody , hydrolysis , protease , serine , biochemistry , amyloid (mycology) , enzyme , microbiology and biotechnology , immunology , biology , inorganic chemistry
gamma secretase results in the production of amyloid beta peptides (A ) that accumulate to form amyloid plaques in Alzheimer’s disease (AD). This process may be regulated in part by APP dimerization as mutants that stabilize dimerization increase A production while mutants that impair dimerization decrease A production. Objective(s): To design and implement the techniques of Bimolecular Fluorescent Complementation (BiFC) and Enzyme Fragment Complementation (EFC) to detect APP dimerization in cells. Both the techniques of BiFC and EFC can be utilized to study the sub-localization of APP dimerization and processing in cells. In addition, we plan to utilize the EFC technique for a high throughput screen of small compounds that inhibit APP dimerization leading to A formation. Methods: In BiFC, inactive fragments of the yellow fluorescent protein (YFP) are fused to the C-terminal end of APP. Upon APP dimerization, the YFP fragments reconstitute into a fully functional fluorescent protein that can be easily detected. Similarly, in EFC, inactive fragments of betagalactosidase ( -gal) are used, and APP dimerization is detected from -gal activity. Results: BiFC has successfully detected APP dimerization qualitatively with immunofluorescence and quantitatively with FACS. Similarly, EFC has successfully detected APP dimerization qualitatively with histochemical staining of -gal and quantitatively with chemiluminescence using a dioxetane compound as substrate. Conclusions: Since it has been shown that APP dimerization is necessary for the toxic A peptide formation, our screen may eventually lead to the identification of APP dimerization inhibitors as a new therapeutic treatment for decreasing A levels in AD.