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Transmembrane Domain Oligomerization Propensity determined by ToxR Assay
JoVE 杂志
生物学
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JoVE 杂志 生物学
Transmembrane Domain Oligomerization Propensity determined by ToxR Assay

Transmembrane Domain Oligomerization Propensity determined by ToxR Assay

DOI:
10.3791/2721-v

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06:45 min

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May 26, 2011

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, ,
1Department of Chemistry and Biochemistry,University of Colorado at Boulder

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

Summary

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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

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