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High-Pressure NMR Experiments for Detecting Protein Low-Lying Conformational States
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JoVE Journal Biochimica
High-Pressure NMR Experiments for Detecting Protein Low-Lying Conformational States

High-Pressure NMR Experiments for Detecting Protein Low-Lying Conformational States

DOI:
10.3791/62701-v

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04:37 min

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June 29, 2021

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, ,
1Department of Chemistry,Iowa State University, 2Roy J. Carver Department of Biochemistry, Biophysics and Molecular Biology,Iowa State University

Capitoli

  • 00:05Introduction
  • 00:34Sample Preparation and High-Pressure Cell Assembly
  • 02:03Recording High-Pressure NMR Experiments
  • 02:32Results: Unfolding of hnRNPA1 RRM2 Domain Under Pressure
  • 04:11Conclusion

Summary

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We provide a detailed description of the steps required to assemble a high-pressure cell, set up and record high-pressure NMR experiments, and finally analyze both peak intensity and chemical shift changes under pressure. These experiments can provide valuable insights into the folding pathways and structural stability of proteins.

Tags

High-pressure NMRProtein Conformational StatesPressure PerturbationGlass CavitiesVoid VolumesNitrogen-15-labeled SampleMineral OilTransmission LiquidPressure Cell AssemblyTransverse Relaxation Optimized SpectroscopyHetero Single-quantum Coherence SpectroscopyFolding/unfolding RatesRRM2Heterogeneous Nuclear Ribonucleoprotein A1

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