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Analyzing Protein Architectures and Protein-Ligand Complexes by Integrative Structural Mass Spectrometry
Published on: October 15, 2018
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Structural Characterization of Native Proteins and Protein Complexes by Electron Ionization Dissociation-Mass
Huilin Li1, Yuewei Sheng2, William McGee3
1Department of Biological Chemistry, David Geffen School of Medicine, University of California , Los Angeles, California 90095, United States.
Analytical Chemistry
|February 15, 2017
Summary
A new mass spectrometry technique, 30 eV electron ionization dissociation (EID), offers enhanced structural insights into large protein complexes. EID provides rich data surpassing electron capture dissociation (ECD) for native protein studies.
Area of Science:
- Proteomics
- Biophysical Chemistry
- Mass Spectrometry
Background:
- Mass spectrometry (MS) is crucial for protein identification and characterization.
- Tandem MS provides versatile structural information for proteins and complexes.
- Electron capture dissociation (ECD) preserves noncovalent interactions but struggles with large proteins due to fragmentation challenges.
Purpose of the Study:
- To introduce and evaluate 30 eV electron ionization dissociation (EID) as a novel activation method for top-down MS.
- To explore EID's capability in characterizing native protein-ligand and protein-protein complexes.
- To compare EID with ECD and ultraviolet photodissociation (UVPD) for native protein analysis.
Main Methods:
- Utilized 30 eV electron ionization dissociation (EID) for top-down mass spectrometry.
- Studied human carbonic anhydrase I (HCA-I) for gas-phase and solution-phase stability.
- Analyzed apo-, Zn-, and Cu,Zn-superoxide dismutase (SOD1) dimers, including a G41D mutation.
- Compared EID performance with electron capture dissociation (ECD) and 193 nm ultraviolet photodissociation (UVPD).
Main Results:
- EID generated rich structural information, exceeding ECD capabilities.
- EID enabled comparison of HCA-I's structural stability and unfolding in gas and solution phases.
- EID fragmentation patterns revealed structural similarities and differences in SOD1 dimers.
- EID-MS identified structural changes in SOD1 due to copper binding and a G41D mutation.
Conclusions:
- 30 eV EID is a powerful tool for top-down MS characterization of native protein complexes.
- EID provides valuable structural insights complementary to ECD and UVPD.
- EID facilitates detailed analysis of protein structure, stability, and ligand interactions.
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