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P1‐229: A MATHEMATICAL MODEL OF ALZHEIMER'S DISEASE SIMULATES DIFFERENT GENE THERAPY TREATMENTS AND THEIR EFFECTS ON MITOCHONDRIAL ACTIVITY
Author(s) -
Coleman Randolph A.,
Shelton Morgan,
Shirali Sonali,
Castora Frank J.
Publication year - 2018
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.2018.06.234
Subject(s) - gene knockdown , mitochondrial biogenesis , gene expression , gene , biology , mitochondrion , genetics , computational biology , microbiology and biotechnology
APP family proteins. The C-terminal region of Nipsnap1 is evolutionarily conserved from humans to bacteria. The molecular and cellular functions of Nipsnap1 remain unknown.Methods:Ahomology model of C-terminal region of mouse Nipsnap1 was generated using bacterial Nipsnap proteins NP_356412.1 (PDB ID 1VQS) and NP_396154.1 (PDB ID 1VQY), and virtual ligand screening was performed using FINDSITE. Recombinant Nipsnap1 proteins (full-length, C-terminal fragment, and N-terminal fragment) were affinity purified from bacteria and mammalian cells. The interaction between recombinant Nipsnap1 proteins and APP cytoplasmic peptide (APP-C18), NAD(H), or NADP(H) were analyzed using biochemical pulldown assays and circular dichroism spectroscopy. Results: FINDSITE virtual ligand screening of over 70,000 compounds identified NADH and NADPH as the top ligand candidates for Nipsnap1 C-terminal region. Using biochemical pull-down assays, we show that purified recombinant Nipsnap1 binds directly to NADH and NADPH. In addition, Nipsnap1 interaction with NADH and NADPH was validated using near-UV circular dichroism spectroscopy. Current experiments are focused on mapping the interaction domain of Nipsnap1 with APP and NADH, and determining if APP interferes with Nipsnap1-NADH binding. Conclusions:The Cterminal region of APP family proteins interacts with mitochondrial Nipsnap1, which also directly binds NAD(P)H. These results suggest that APP C-terminal fragment may regulate NAD(P)H metabolism through an interaction with Nipnsap1.