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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
Eine neuartige Methode, die quantitative Analyse von FRET (Förster Resonance Energy Transfer)-Signale für das Studium Enzymkinetik beschrieben.<em> K<sub> M</sub</em> Und<em> K<sub> Cat</sub</em> Wurden zur Hydrolyse der katalytischen Domäne des SENP1 (SUMO / Sentrin spezifische Protease 1) vor SUMO1 (Small Ubiquitin-ähnlicher Modifikator) erhalten. Die allgemeinen Grundsätze dieses quantitative-FRET-basierten Protease kinetische Untersuchung auf andere Proteasen angewendet werden.
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
