Journal
MOLECULAR NEURODEGENERATION
Volume 4, Issue -, Pages -Publisher
BMC
DOI: 10.1186/1750-1326-4-22
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Funding
- NIH NRSA predoctoral fellowship [5F31NS053218-02, AG026660]
- Alzheimer's Association
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gamma-Secretase is an aspartyl protease that cleaves multiple substrates that are involved in broad biological processes ranging from stem cell development to neurodegeneration. The investigation of gamma-secretase has been limited by currently available assays that require genetic or biochemical manipulation in the form of substrate transfection or membrane preparation. Here we report an exo-cell assay that is capable of characterizing gamma-secretase activity in any cellular system without limitation. Using a highly active, recombinant substrate this assay can quickly and easily ascertain the status of gamma-secretase activity in cell systems and patient samples. We have applied this method to determine the activity of gamma-secretase in primary cell samples where transfection and/or membrane isolation are not viable options. Importantly, it allows for the detection of real time gamma-secretase activity after inhibitor or drug treatment. The application of this assay to determine the role of gamma-secretase in physiological and pathological conditions will greatly facilitate our characterization of this complex protease and help in the development and evaluation of gamma-secretase targeted therapies in Alzheimer's disease or a variety of neoplasms.
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