This content is Free Access.
Chapters
- 00:05Title
- 01:32Protein Expression Cysteine Mutation and mRNA Synthesis
- 02:07Oocyte Removal
- 03:36Oocyte Defoculation and mRNA Injection
- 05:05Site-specific Fluorescence Labeling
- 06:07Electrophysiology
- 07:26Anisotropy Measurements and Determination of Distance Constraints
- 09:17Relative Movement of Protein Subunits
- 10:07Results: Current and Fluorescence Changes with Site Specific Fluorophore Labelings
- 11:38Conclusion
Ci descrive un metodo che misura la cinetica del trasporto ionico di proteine di membrana a fianco sito-specifica analisi dei cambiamenti conformazionali mediante fluorescenza su singole cellule. Questa tecnica è adattabile a canali ionici, trasportatori e pompe ioniche e può essere utilizzato per determinare vincoli di distanza tra le subunità proteiche.
Tags
Conformational DynamicsMembrane ProteinsSite-directed Fluorescence LabelingTwo Electrode Voltage Clamp Electrophysiology (TEVC)Ion TransportIon ChannelsIon PumpsTransportersSite-specific Fluorophore LabelingConformational DynamicsVoltage-clamp FluorometryCysteine ReplacementSite-directed Fluorophore LabelingMolecular MotionDistance ConstraintsXenopus Leavis OocytesFunctional ExpressionFluorescence Signal
