Journal
/
Chemistry
/
Monitoring Protein-Ligand Interactions in Human Cells by Real-Time Quantitative In-Cell NMR using a High Cell Density Bioreactor
JoVE Journal
Chemistry
This content is Free Access.
JoVE Journal Chemistry
Monitoring Protein-Ligand Interactions in Human Cells by Real-Time Quantitative In-Cell NMR using a High Cell Density Bioreactor

Monitoring Protein-Ligand Interactions in Human Cells by Real-Time Quantitative In-Cell NMR using a High Cell Density Bioreactor

DOI:
10.3791/62323-v

•

10:25 min

•

March 09, 2021

•

,
1Magnetic Resonance Center − CERM,University of Florence, 2Consorzio Interuniversitario Risonanze Magnetiche di Metalloproteine − CIRMMP, 3Neurofarba Department,University of Florence

Chapters

  • 00:05Introduction
  • 00:47Bioreactor Setup
  • 01:34Preparation of the Cell Sample
  • 04:03Bioreactor Operation and NMR Experiments
  • 06:00MCR-ALS Analysis
  • 08:06Results: Monitoring Protein-Ligand Interactions by Real-Time Quantitative In-Cell NMR
  • 09:48Conclusion

Summary

Automatic Translation

Automatic Translation

This protocol describes the setup of an NMR bioreactor to keep encapsulated human cells viable for up to 72 h, followed by time-resolved in-cell NMR data acquisition and analysis. The methodology is applied to monitor intracellular protein-ligand interactions in real time.

Tags

Keywords: In-cell NMRProtein-ligand InteractionsHuman CellsBioreactorReal-time MonitoringProtein StructureProtein FunctionConformational ChangesChemical ChangesIsotope LabelingHEK 293T CellsAgarose ThreadsCell EmbeddingFlow UnitNMR TubeTemperature ControlCell ViabilityCell Metabolism

Article

Embed

ADD TO PLAYLIST

Usage Statistics

Related Videos

check_url/62323?article_type=v

Untargeted Metabolomics from Biological Sources Using Ultraperformance Liquid Chromatography-High Resolution Mass Spectrometry (UPLC-HRMS)
check_url/62323?article_type=v

Quantitative and Qualitative Examination of Particle-particle Interactions Using Colloidal Probe Nanoscopy
check_url/62323?article_type=v

Real-time Monitoring of Reactions Performed Using Continuous-flow Processing: The Preparation of 3-Acetylcoumarin as an Example
check_url/62323?article_type=v

Radiolabeling and Quantification of Cellular Levels of Phosphoinositides by High Performance Liquid Chromatography-coupled Flow Scintillation
check_url/62323?article_type=v

Dissolution Dynamic Nuclear Polarization Instrumentation for Real-time Enzymatic Reaction Rate Measurements by NMR
check_url/62323?article_type=v

Flow-pattern Guided Fabrication of High-density Barcode Antibody Microarray
check_url/62323?article_type=v

Rapid High-throughput Species Identification of Botanical Material Using Direct Analysis in Real Time High Resolution Mass Spectrometry
check_url/62323?article_type=v

Spin Saturation Transfer Difference NMR (SSTD NMR): A New Tool to Obtain Kinetic Parameters of Chemical Exchange Processes
check_url/62323?article_type=v

Capillary Electrophoresis to Monitor Peptide Grafting onto Chitosan Films in Real Time
check_url/62323?article_type=v

Real-time Breath Analysis by Using Secondary Nanoelectrospray Ionization Coupled to High Resolution Mass Spectrometry

Read Article