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Chapters
- 00:05Title
- 01:26Expression of the Chimeric Proteins
- 02:30Measurement of β-Galactosidase Activity
- 03:40Verification of Protein Expression
- 04:24Control for Proper Membrane Insertion
- 05:34Representative Results of the ToxR Transcriptional Reporter Assay
- 06:17Conclusion
An efficient procedure to assess the oligomerization propensity of single-pass transmembrane domains (TMDs) is described. Chimeric proteins consisting of the TMD fused to ToxR are expressed in an E. coli reporter strain. TMD-induced oligomerization causes dimerization of ToxR, activation of transcription and production of the reporter protein, -galactosidase.
Tags
Transmembrane DomainOligomerization PropensityToxR AssayProtein Transmembrane DomainsPhospholipid BilayersMembrane-spanning ProteinsHydrophobic HelicesStructured Transmembrane OligomersAmino AcidsMembrane EnvironmentAqueous SolutionIntermolecular ForcesProtein AssociationMolecular RecognitionWater-soluble RegionsTransmembrane OligomerizationAssociation MeasurementIntegral Membrane Protein FunctionSynthetic PeptidesCellular Membranes
