b- secretase (BACE1) is a key protease involved in the production of beta-amyloid peptides found in extracellular amyloid plaques of patients with Alzheimer’s disease (AD).1 This enzyme has been implicated as an important target for anti-amyloid therapy of AD.2
The recombinant human BACE1 contains the catalytic domain and a C-terminal His tag. The protein was expressed as inclusion bodies in E. coli. It was refolded and purified using ion exchange and IMAC chromatography.3 Enzyme activity can been measured in fluorescence resonance energy transfer (FRET) substrate based assays (e.g. SensoLyte® 520 beta-Secretase Assay Kit, cat# 71144). 100 ng of b-secretase is sufficient for this FRET-based assay.
AnaSpec’s SensoLyte™ 520 b-Secretase Assay Kit (cat# 71144) is the industry’s only long wavelength assay kit in the green range. This kit is a fluorimetric assay kit and is sensitive, robust and easy-to-use. It employs a FRET peptide (mentioned above), facilitating high throughput screening of beta-secretase targeting drug candidates. The sequence of this FRET peptide is derived from the beta-secretase cleavage site of beta-amyloid precursor protein (APP) with Swedish mutation. This mutation enhances the susceptibility of APP to beta-secretase and results in an early onset of AD.
References:
1. Willem, M. et al. Semin Cell Dev Biol. 20,175 (2009).
2. Ghosh, K. et al. Neurotherapeutics 5, 399 (2008).
3. Tomasselli, G. et al. Protein & Peptide Letters. 15,131 (2008).
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