Journal
/
Biochemistry
/
Analyzing Dynamic Protein Complexes Assembled On and Released From Biolayer Interferometry Biosensor Using Mass Spectrometry and Electron Microscopy
JoVE Journal
Biochemistry
This content is Free Access.
JoVE Journal Biochemistry
Analyzing Dynamic Protein Complexes Assembled On and Released From Biolayer Interferometry Biosensor Using Mass Spectrometry and Electron Microscopy

Analyzing Dynamic Protein Complexes Assembled On and Released From Biolayer Interferometry Biosensor Using Mass Spectrometry and Electron Microscopy

DOI:
10.3791/57902-v

•

09:30 min

•

August 06, 2018

•

, , , , ,
1Department of Biochemistry and Molecular Biology,University of Kansas Medical Center, 2Department of Chemistry,Massachusetts Institute of Technology

Chapters

  • 00:04Title
  • 01:28Assembly of Prepore Anthrax Toxin Complex on a PDEA-modified Amine Reactive Biosensor Surface
  • 04:37Assembly of Pore Anthrax Toxin Complex on PDEA-modified Amine Reactive Biosensor Surface
  • 05:26Identification of Released Macromolecular Assemblies from BLI Biosensors by Negative Stain Electron Microscopy and Mass Spectrometry
  • 06:50Results: Analysis of Dynamic Protein Complexes by Biolayer Interferometry, Electron Microscopy, and Mass Spectrometry
  • 08:29Conclusion

Summary

Automatic Translation

Automatic Translation

Here we present a protocol to monitor the assembly and disassembly of the anthrax toxin using biolayer interferometry (BLI). Following assembly/disassembly on the biosensor surface, the large protein complexes are released from the surface for visualization and identification of components of the complexes using electron microscopy and mass spectrometry, respectively.

Tags

Keywords: Protein ComplexesBiolayer InterferometryMass SpectrometryElectron MicroscopyAnthrax ToxinBiosensorProtein AssemblyPH-induced Structural RearrangementsLFnNHSEDCPDEAL-cysteine

Article

Embed

ADD TO PLAYLIST

Usage Statistics

Related Videos

check_url/57902?article_type=v

High-resolution Single Particle Analysis from Electron Cryo-microscopy Images Using SPHIRE
check_url/57902?article_type=v

Time-resolved ElectroSpray Ionization Hydrogen-deuterium Exchange Mass Spectrometry for Studying Protein Structure and Dynamics
check_url/57902?article_type=v

Resolving Affinity Purified Protein Complexes by Blue Native PAGE and Protein Correlation Profiling
check_url/57902?article_type=v

Analysis of β-Amyloid-induced Abnormalities on Fibrin Clot Structure by Spectroscopy and Scanning Electron Microscopy
check_url/57902?article_type=v

Detection of Protein Ubiquitination Sites by Peptide Enrichment and Mass Spectrometry
check_url/57902?article_type=v

Application of Biolayer Interferometry (BLI) for Studying Protein-Protein Interactions in Transcription
check_url/57902?article_type=v

Quantitative Analysis of the Cellular Lipidome of Saccharomyces Cerevisiae Using Liquid Chromatography Coupled with Tandem Mass Spectrometry
check_url/57902?article_type=v

Semi-Quantitative Analysis of Peptidoglycan by Liquid Chromatography Mass Spectrometry and Bioinformatics
check_url/57902?article_type=v

Quantitative Metabolomics of Saccharomyces Cerevisiae Using Liquid Chromatography Coupled with Tandem Mass Spectrometry
check_url/57902?article_type=v

A Mass Spectrometry-Based Proteomics Approach for Global and High-Confidence Protein R-Methylation Analysis

Read Article