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Measuring human β-secretase (BACE1) activity using homogeneous time-resolved fluorescence by Matthew E. Kennedy; Wenyan Wang; Lixin Song; Julie Lee; Lili Zhang; Gwen Wong; Liyang Wang; Eric Parker is a Biochemistry, Genetics and Molecular Biology article available to read on EtoBox.
What is Measuring human β-secretase (BACE1) activity using homogeneous time-resolved fluorescence about?
The human beta-secretase enzyme, BACE1, mediates a critical step in the production of A beta(40) and A beta(42) peptides which are responsible for the severe neuronal cell death and insoluble amyloid plaques of Alzheimer's disease (AD). Several lines of evidence suggest that potent BACE1 inhibitors represent an attractive A beta-lowering strategy for AD. We designed a simple homogeneous time-resolved fluorescence (HTRF) assay which utilizes the fluorescence resonance energy transfer (FRET) pair europium and allophycocyanin for measuring BACE1 enzymatic activity in a high-throughput manner. Robust FRET was observed when an 18-amino-acid APP Swedish-synthetic peptide that was N-terminally labeled with europium cryptate and C-terminally biotinylated was incubated with streptavidin-coupled cross-linked allophycocyanin (SA-XL665). Purified BACE1 enzyme caused a time- and concentration-dependent linear change in FRET at low nanomolar enzyme concentrations. This assay was used to compare the autoprocessed "mature" BACE1 enzyme (sautoBACe1) and the soluble proBACE1 for activity and inhibition by selected peptidic BACE inhibitors. sautoBACE1 displayed only a modest increase in activity comp
Who reads Measuring human β-secretase (BACE1) activity using homogeneous time-resolved fluorescence?
It is typically read by researchers, students, and practitioners in Biochemistry, Genetics and Molecular Biology.
- Author
- Matthew E. Kennedy; Wenyan Wang; Lixin Song; Julie Lee; Lili Zhang; Gwen Wong; Liyang Wang; Eric Parker
- Publisher
- Elsevier Science; Elsevier ; Elsevier Inc.; Elsevier BV (ISSN 0003-2697)
- Published
- 2003
- Language
- EN
- Field
- Biochemistry, Genetics and Molecular Biology (Life Sciences)