Resolving heterogeneous macromolecular assemblies by Orbitrap-based single-particle charge detection mass
Tobias P Wörner1,2, Joost Snijder1,2, Antonette Bennett3
1Biomolecular Mass Spectrometry and Proteomics, Bijvoet Center for Biomolecular Research and Utrecht Institute for Pharmaceutical Sciences, University of Utrecht, Utrecht, the Netherlands.
Nature Methods
|March 11, 2020
Summary
Single-particle charge detection mass spectrometry on an Orbitrap analyzes large biomolecular assemblies. This method resolves mixed ion populations and determines ion masses, enabling ultrasensitive analysis of complex protein structures.
Area of Science:
- Biophysical techniques
- Mass spectrometry
- Molecular biology
Background:
- Megadalton biomolecular assemblies present analytical challenges.
- Characterizing heterogeneous protein complexes requires sensitive methods.
Purpose of the Study:
- To demonstrate single-particle charge detection mass spectrometry on an Orbitrap.
- To enable ultrasensitive analysis of megadalton biomolecular assemblies.
Main Methods:
- Utilizing Orbitrap mass spectrometry for single-particle analysis.
- Establishing linear scaling of signal amplitude with ion charge.
- Applying charge detection to resolve ion populations and determine masses.
Main Results:
- Demonstrated linear signal amplitude-charge relationship for individual ions.
- Successfully resolved mixed ion populations and determined charge states.
- Enabled mass determination for individual ions within heterogeneous assemblies.
Conclusions:
- Single-particle charge detection mass spectrometry on Orbitrap is effective for megadalton assemblies.
- The method allows for ultrasensitive characterization of complex and heterogeneous biomolecular structures.
- This technique advances the analysis of large protein complexes like ribosomes and viruses.
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