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Millisecond Hydrogen/Deuterium-Exchange Mass Spectrometry for the Study of Alpha-Synuclein Structural Dynamics Under Physiological Conditions
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JoVE Journal Neuroscienze
Millisecond Hydrogen/Deuterium-Exchange Mass Spectrometry for the Study of Alpha-Synuclein Structural Dynamics Under Physiological Conditions

Millisecond Hydrogen/Deuterium-Exchange Mass Spectrometry for the Study of Alpha-Synuclein Structural Dynamics Under Physiological Conditions

DOI:
10.3791/64050-v

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08:40 min

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June 23, 2022

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,
1Living Systems Institute, Department of Biosciences,University of Exeter

Capitoli

  • 00:04Introduction
  • 00:41Peptide Mapping Procedure
  • 02:38Millisecond Hydrogen/Deuterium Exchange Study
  • 05:34Data Processing and Data Analysis
  • 07:19Results: Measuring the Conformational Dynamics of aSyn
  • 08:14Conclusion

Summary

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The structural ensemble of monomeric alpha-synuclein affects its physiological function and physicochemical properties. The present protocol describes how to perform millisecond hydrogen/deuterium-exchange mass spectrometry and subsequent data analyses to determine conformational information on the monomer of this intrinsically disordered protein under physiological conditions.

Tags

Alpha-synucleinHydrogen/deuterium ExchangeMass SpectrometryIntrinsically Disordered ProteinsPeptide MappingSample PreparationAutosamplerHDX RoboticsQuench BufferPeptide Coverage MapFast HDX Prototype InstrumentHDX Data Analysis Software

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