Trioxane-based MS-cleavable Cross-linking Mass Spectrometry for Profiling Multimeric Interactions of Cellular
Biorxiv : the Preprint Server for Biology
|August 30, 2024
Summary
A new cross-linking reagent, TSTO, enables precise mapping of protein-protein interactions (PPIs) by identifying trimeric connections. This advances structural systems biology by revealing complex structural details from living cells.
Area of Science:
- Biochemistry
- Structural Biology
- Proteomics
Background:
- Cross-linking mass spectrometry (XL-MS) maps protein-protein interactions (PPIs) using distance restraints.
- Multimeric cross-links offer higher spatial resolution for complex protein structures but are difficult to identify.
- Existing methods face challenges in identifying multimeric cross-links due to fragmentation complexity and search space expansion.
Purpose of the Study:
- To develop a novel cross-linking reagent for efficient identification of multimeric cross-links.
- To enhance the spatial resolution and accuracy of XL-MS for studying protein complexes.
- To advance the field of structural systems biology through improved PPI mapping.
Main Methods:
- Development of a novel trioxane-based, MS-cleavable homotrifunctional cross-linker, TSTO.
- Simultaneous targeting of three proximal lysine residues by TSTO.
- Utilizing LC-MS^n analysis for fast and unambiguous identification of TSTO-cross-linked peptides.
Main Results:
- TSTO enables rapid and unambiguous identification of cross-linked peptides.
- The TSTO-based XL-MS platform effectively maps PPIs in protein complexes and cellular networks.
- Trimeric interactions captured by TSTO reveal novel structural details missed by existing reagents.
Conclusions:
- TSTO significantly advances XL-MS technology for global PPI profiling from living cells.
- The development of TSTO provides a new direction for creating multifunctional MS-cleavable cross-linkers.
- This work facilitates in-depth PPI description and structural modeling, pushing structural systems biology forward.
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