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Chapters
- 00:05Title
- 01:10Y2H: Yeast Transformation
- 03:22Y2H: Colony-lift Filter Paper β-Gal Assay
- 04:29Protein Expression in a Mammalian Cell Line
- 06:56Co-immunoprecipitation
- 09:02Chemical Cross-linking
- 09:46Results: The Ryanodine Receptor Forms Homo-oligomers Within Cells
- 11:40Conclusion
Oligomerization of the ryanodine receptor, a homo-tetrameric ion channel mediating Ca2+ release from intracellular stores, is critical for skeletal and cardiac muscle contraction. Here, we present complementary in vivo and in vitro methods to detect protein self-association and determine homo-oligomer stoichiometry.
Tags
Protein OligomerizationRyanodine ReceptorYeast Two-hybridCo-immunoprecipitationChemical Cross-linkingProtein-protein InteractionsHomo-oligomersCardiopathiesCardiac ArrhythmiasSudden DeathAnti-arrhythmic DrugsCompetent Yeast CellsPlasmid DNALithium AcetatePEG SolutionDimethyl SulfoxideMinimal SD Medium Plates
