The primary roles of Core B include the quantification of AS levels by ELISAs, the development of novel ELISA and y-secretase assays, and the maintenance and distribution of the Program's extensive reagents. First, Core B will continue to perform AS ELISAs for monomeric total AS, A61-40, ASx-40, ASx-42 and AS1- 42 for all investigators involved in the Program. We will measure the concentration of monomeric AS in conditioned media from stable cell lines and primary neuronal cultures, cell lysates, subcellular fractions, mouse and human brain homogenates, and transgenic mouse plasma. The Core will provide an effective and convenient means for obtaining quantitative measurements of AS for all four projects and the animal core. Second, Core B will develop a specific ELISA for measuring soluble oligomeric AS species. Low molecular weight AS oligomers play significant roles in neuronal metabolism, and the levels of soluble AS dimers/trimers/tetramers will be measured by our newly developed ELISA. Third, Core B will establish new ELISA methods to quantify the y-secretase cleavage of Notch and APLP2. Levels of AS-like peptides will be measured by ELISA by quantifying the y-secretase cleavages of chimeric APP-Notch and APP-APLP2 proteins. Fourth, Core B will perform standard y-secretase activity assay and develop new approaches to modulate the y-secretase activity in vitro. Further, Core B will carry out the in vitro y-secretase assay under new conditions proposed by a Program Investigator. Finally, Core B will continue to be responsible for maintaining and supplying the extensive shared cell lines, DMA constructs and antibodies of the Program to all of our investigators as well as to outside investigators, as in the past. Core B thus serves as a vital central repository for a large number of highly useful reagents that have been generated and characterized over the past two decades by the Investigators involved in this Program Project. These reagents include hundreds of cDNA constructs, several dozen stably-transfected mammalian cell lines and an extensive collection of specific polyclonal and monoclonal antibodies. The mission of Core B is to greatly facilitate ongoing collaborations and overall experimental efficiency within the Program Project.

Agency
National Institute of Health (NIH)
Institute
National Institute on Aging (NIA)
Type
Research Program Projects (P01)
Project #
5P01AG015379-15
Application #
8381465
Study Section
Special Emphasis Panel (ZAG1-ZIJ-3)
Project Start
Project End
2013-08-31
Budget Start
2012-09-01
Budget End
2013-08-31
Support Year
15
Fiscal Year
2012
Total Cost
$123,277
Indirect Cost
$76,736
Name
Brigham and Women's Hospital
Department
Type
DUNS #
030811269
City
Boston
State
MA
Country
United States
Zip Code
02115
Zoltowska, Katarzyna Marta; Berezovska, Oksana (2017) Dynamic Nature of presenilin1/?-Secretase: Implication for Alzheimer's Disease Pathogenesis. Mol Neurobiol :
Gong, Yi; Sasidharan, Nikhil; Laheji, Fiza et al. (2017) Microglial dysfunction as a key pathological change in adrenomyeloneuropathy. Ann Neurol 82:813-827
Kara, Eleanna; Marks, Jordan D; Fan, Zhanyun et al. (2017) Isoform- and cell type-specific structure of apolipoprotein E lipoparticles as revealed by a novel Forster resonance energy transfer assay. J Biol Chem 292:14720-14729
Raven, Frank; Ward, Joseph F; Zoltowska, Katarzyna M et al. (2017) Soluble Gamma-secretase Modulators Attenuate Alzheimer's ?-amyloid Pathology and Induce Conformational Changes in Presenilin 1. EBioMedicine 24:93-101
Wagner, Steven L; Rynearson, Kevin D; Duddy, Steven K et al. (2017) Pharmacological and Toxicological Properties of the Potent Oral ?-Secretase Modulator BPN-15606. J Pharmacol Exp Ther 362:31-44
Yang, Ting; Li, Shaomin; Xu, Huixin et al. (2017) Large Soluble Oligomers of Amyloid ?-Protein from Alzheimer Brain Are Far Less Neuroactive Than the Smaller Oligomers to Which They Dissociate. J Neurosci 37:152-163
Ward, Joseph; Wang, Haizhi; Saunders, Aleister J et al. (2017) Mechanisms that synergistically regulate ?-secretase processing of APP and A?-? protein levels: relevance to pathogenesis and treatment of Alzheimer's disease. Discov Med 23:121-128
Zoltowska, Katarzyna Marta; Maesako, Masato; Lushnikova, Iryna et al. (2017) Dynamic presenilin 1 and synaptotagmin 1 interaction modulates exocytosis and amyloid ? production. Mol Neurodegener 12:15
Bolduc, D M; Selkoe, D J; Wolfe, M S (2017) Enzymatic Assays for Studying Intramembrane Proteolysis. Methods Enzymol 584:295-308
Williamson, Rebecca L; Laulagnier, Karine; Miranda, André M et al. (2017) Disruption of amyloid precursor protein ubiquitination selectively increases amyloid ? (A?) 40 levels via presenilin 2-mediated cleavage. J Biol Chem 292:19873-19889

Showing the most recent 10 out of 139 publications