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Approches génétiques et biochimiques pour<em> In Vivo</em> et<em> In vitro</em> Évaluation de Protein oligomérisation: Étude de cas Ryanodine Receptor
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Biology
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JoVE Journal Biology
Genetic and Biochemical Approaches for In Vivo and In Vitro Assessment of Protein Oligomerization: The Ryanodine Receptor Case Study
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Approches génétiques et biochimiques pour<em> In Vivo</em> et<em> In vitro</em> Évaluation de Protein oligomérisation: Étude de cas Ryanodine Receptor

DOI:
10.3791/54271-v

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12:43 min

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July 27, 2016

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, ,
1Wales Heart Research Institute,Cardiff University School of Medicine, College of Biomedical and Life Sciences

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

Summary

Automatic Translation

Automatic Translation

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

Keywords: Protein OligomerizationRyanodine ReceptorYeast Two-hybridCo-immunoprecipitationChemical Cross-linkingProtein-protein InteractionsHomo-oligomersCardiopathiesCardiac ArrhythmiasSudden DeathAnti-arrhythmic DrugsCompetent Yeast CellsPlasmid DNALithium AcetatePEG SolutionDimethyl SulfoxideMinimal SD Medium Plates

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