An exo-cell assay for examining real-time γ-secretase activity and inhibition Journal Article
| Authors: | Shelton, C. C.; Tian, Y.; Frattini, M. G.; Li, Y. M. |
| Article Title: | An exo-cell assay for examining real-time γ-secretase activity and inhibition |
| Abstract: | 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 -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 -secretase activity in any cellular system without limitation. Using a highly active, recombinant substrate this assay can quickly and easily ascertain the status of -secretase activity in cell systems and patient samples. We have applied this method to determine the activity of -secretase in primary cell samples where transfection and/or membrane isolation are not viable options. Importantly, it allows for the detection of real time -secretase activity after inhibitor or drug treatment. The application of this assay to determine the role of -secretase in physiological and pathological conditions will greatly facilitate our characterization of this complex protease and help in the development and evaluation of -secretase-targeted therapies in Alzheimer's disease or a variety of neoplasms. © 2009 Shelton et al; licensee BioMed Central Ltd. |
| Keywords: | controlled study; unclassified drug; human cell; nonhuman; drug targeting; neoplasm; mass spectrometry; animal cell; mouse; enzyme inhibition; drug potency; drug structure; enzyme activity; enzyme substrate; hela cell; cell assay; stem cell; dna; genetic transfection; neuroblastoma cell; protein purification; lymphoma cell; recombinant protein; chronic lymphatic leukemia; liquid chromatography; cell activity; enzyme assay; alzheimer disease; peripheral blood mononuclear cell; amyloid precursor protein; gamma secretase inhibitor; gsi 34; thrombin; benzodiazepine; gamma secretase; biotinylation; biotin; aspartic proteinase; dna fragment; l 685458; tritium; b cell leukemia; exo cell assay; nerve degeneration |
| Journal Title: | Molecular Neurodegeneration |
| Volume: | 4 |
| Issue: | 1 |
| ISSN: | 1750-1326 |
| Publisher: | Biomed Central Ltd |
| Date Published: | 2009-06-02 |
| Start Page: | 22 |
| Language: | English |
| DOI: | 10.1186/1750-1326-4-22 |
| PROVIDER: | scopus |
| PUBMED: | 19490610 |
| PMCID: | PMC3224971 |
| DOI/URL: | |
| Notes: | --- - "Export Date: 30 November 2010" - "Art. No.: 22" - "Source: Scopus" |
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