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P1‐063: THE PPARGAMMA COFACTOR RIP140 REGULATES BACE1 GENE EXPRESSION
Author(s) -
Blondrath Katrin,
Sastre Magdalena
Publication year - 2014
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.2014.05.299
Subject(s) - gene knockdown , chemistry , immunostaining , antibody , microbiology and biotechnology , receptor , biochemistry , biology , gene , immunohistochemistry , immunology
Hydrogen/deuterium exchange (HDX) Mass Spectrometry (MS) analysis to verify the structural changes in A b 40 that occur during incorporation into the lipid membranes. Results: In our experiments, we determined appropriate lipid composition for Ab40 interaction with liposomes. Firstly, we found that Ab40 exhibited the highest affinity to negatively charged ganglioside GM1 among various lipids (Fig. 1A). In the subsequent experiments we established proper lipids composition in large unilamellar vesicles. The western-blot analysis showed the most intensive interaction of Ab with liposomes containing DPPC/GM1/cholesterol (molar ratio 5:2:3) (Fig. 1B). Afterwards calcium uptake experiments revealed that incubation of Ab40 in the presence of liposomes leads to Ca2+ influx into to the vesicles, in a concentration dependent manner (Fig. 2).In the structural analysis of Ab40 peptide during interactions with lipids bilayer using HDXMS we observed that the interaction with liposomes causes changes in the middle part of the peptide (residues from 10 thto 35 thamino acid). This region is protected against hydrogen/deuterium exchange what suggests that this region is less dynamic and more stable and structured (Fig. 3B).Conclusions: These experiments suggests that the middle part of Ab40 is involved in lipid-protein interactions and could participate in the ion channel formation what may play a significant and inevitable cytotoxic role in the pathogenesis of AD.