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Bio-layer Interferometry for Measuring Kinetics of Protein-protein Interactions and Allosteric Ligand Effects
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JoVE Journal Chimica
Bio-layer Interferometry for Measuring Kinetics of Protein-protein Interactions and Allosteric Ligand Effects

Bio-layer Interferometry for Measuring Kinetics of Protein-protein Interactions and Allosteric Ligand Effects

DOI:
10.3791/51383-v

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13:57 min

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February 18, 2014

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,
1Department of Biochemistry and Molecular Biology,SUNY Upstate Medical University

Capitoli

  • 00:05Titolo
  • 02:05Programming the Instrument for BLI Assay
  • 05:44Sample Preparation
  • 07:52Data Processing and Analysis
  • 10:40Results: Analysis of BLI Kinetics to Determine Rate Constants and Binding Affinity
  • 13:14Conclusion

Summary

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The protocols here describe kinetic assays of protein-protein interactions with Bio-layer Interferometry. F-type ATP synthase, which is involved in cellular energy metabolism, can be inhibited by its ε subunit in bacteria. We have adapted Bio-layer Interferometry to study interactions of the catalytic complex with ε’s inhibitory C-terminal domain.

Tags

Protein-protein InteractionsAllosteric Ligand EffectsBio-layer InterferometryATP SynthaseSubunit εBacterial F-type ATP SynthaseConformational ChangeCatalytic ComplexInhibitory InteractionKineticsDrug Discovery

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