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Laser-free Hydroxyl Radical Protein Footprinting to Perform Higher Order Structural Analysis of Proteins
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生物化学
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JoVE 杂志 生物化学
Laser-free Hydroxyl Radical Protein Footprinting to Perform Higher Order Structural Analysis of Proteins

Laser-free Hydroxyl Radical Protein Footprinting to Perform Higher Order Structural Analysis of Proteins

DOI:
10.3791/61861-v

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09:59 min

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June 04, 2021

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, , ,
1GenNext Technologies Inc., 2Department of Biomolecular Sciences, School of Pharmacy,University of Mississippi, 3Department of Chemistry and Biochemistry,University of Mississippi

Chapters

  • 00:04Introduction
  • 00:35Installing the Capillary Tube
  • 02:43Installing an Injection Loop
  • 03:34Initializing the Laser Free HRPF System
  • 04:45Modification of Protein to Detect Changes in Higher Order Structure
  • 06:59Shutting Down the Laser Free HRPF System
  • 07:43Results: High Order Structural Analysis of Proteins Using Laser-free Hydroxyl Radical Protein Footprinting
  • 09:15Conclusion

Summary

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This protocol presents a method to use inline radical dosimetry and a plasma light source to perform flash oxidation protein footprinting. This method replaces the hazardous UV laser to simplify and improve the reproducibility of fast photochemical oxidation of protein studies.

Tags

Protein FootprintingHydroxyl RadicalLaser-freeProtein StructureProtein InteractionProtein AggregationProtein StabilityCapillaryPhotolysisDosimetrySilicaPolyimideNutFerrulePhotolysis CellDosimeter CellMagnetic MaskTeflon TubingInjection Loop

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