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Kvantitativ FRET (Förster Resonance Energy Transfer) Analyse for SENP1 Protease Kinetics Bestemmelse
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Chapters
- 00:05Title
- 02:20Plasmid Constructs
- 03:07Protein Expression and Purification
- 06:16FRET-based Protease Assay for Enzyme Kinetic Study
- 08:54Quantitative FRET Spectrum Analysis
- 13:36Representative Results
- 15:11Conclusion
En hidtil ukendt fremgangsmåde indebærer kvantitativ analyse af FRET (Förster Resonance Energy Transfer) signaler beskrives til undersøgelse af enzymkinetik.<em> K<sub> M</sub</em> Og<em> K<sub> Kat</sub</em> Opnåedes til hydrolyse af det katalytiske domæne af SENP1 (SUMO / Sentrin specifik protease 1) til præ-SUMO1 (Small ubiquitin-lignende modifier). De generelle principper i denne kvantitative-FRET-baserede protease kinetisk undersøgelse kan anvendes på andre proteaser.
Tags
Quantitative FRET AnalysisF&246;rster Resonance Energy TransferSENP1 Protease Kinetics DeterminationPosttranslational ModificationsUbiquitinUbiquitin-like ProteinsProtein Activity RegulationSUMO ModificationCellular ProcessesSUMO-specific ProteaseEndopeptidaseIsopeptidaseCatalytic EfficiencySpecificityKinetic ConstantsKcat/KM RatioMethods For Determining Kinetic ParametersPolyacrylamide Gel-based Western-blotRadioactive-labeled SubstrateFluorescent CompoundProtein Labeled Substrate
