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¹H NMR of Labile Protons: Deuterium (²H) Substitution00:48

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High-Resolution Mass Spectrometry (HRMS)01:15

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Related Experiment Video

Updated: Jul 5, 2026

Analyzing Protein Dynamics Using Hydrogen Exchange Mass Spectrometry
11:37

Analyzing Protein Dynamics Using Hydrogen Exchange Mass Spectrometry

Published on: November 29, 2013

Dynamic structural changes during complement C3 activation analyzed by hydrogen/deuterium exchange mass spectrometry.

Michael C Schuster1, Daniel Ricklin, Krisztián Papp

  • 1Department of Medicine, Division of Rheumatology, University of Pennsylvania, Philadelphia, PA USA.

Molecular Immunology
|May 6, 2008
PubMed
Summary

Complement component C3 activation to C3b involves dynamic structural changes in solution. Hydrogen/deuterium exchange mass spectrometry (HDX-MS) revealed altered solvent accessibility in C3b, highlighting flexible regions crucial for complement function.

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Area of Science:

  • Immunology
  • Structural Biology
  • Biochemistry

Background:

  • Complement component C3 cleavage to C3b is central to complement activation.
  • C3b is biologically active, mediating various complement functions.
  • Existing crystal structures of C3 and C3b lack dynamic solution-phase information.

Purpose of the Study:

  • To investigate structural changes of C3 and C3b in solution using HDX-MS.
  • To assess the feasibility of HDX-MS for large plasma proteins like C3.
  • To correlate solution structures with known crystal structures.

Main Methods:

  • Hydrogen/deuterium exchange coupled with mass spectrometry (HDX-MS).
  • Utilized two forms of mass spectrometry for enhanced sequence coverage of C3b.
  • Analysis of 82 peptides to track changes in solvent accessibility over time.

Main Results:

  • HDX-MS successfully mapped structural dynamics of C3 and C3b in solution.
  • Observed significant changes in solvent accessibility for 16 peptides in the alpha-chain of C3b.
  • Identified a flexible, discontinuous region in C3b involving MG3, MG6, LNK, and alpha'NT domains.

Conclusions:

  • HDX-MS provides insights into the dynamic structural transitions of C3 and C3b in solution.
  • The identified flexible region in C3b may be critical for complement activation and function.
  • This study demonstrates the utility of HDX-MS for studying large, dynamic protein systems.