Anatomy of a crosslinker
Adam Belsom1, Juri Rappsilber2
1Bioanalytics, Institute of Biotechnology, Technische Universität Berlin, 13355, Berlin, Germany.
Current Opinion in Chemical Biology
|August 24, 2020
Summary
Crosslinking mass spectrometry relies on reagents. This study advocates for data-driven design of crosslinkers, focusing on reaction kinetics and enrichment groups for improved structural biology applications.
Area of Science:
- Structural biology
- Systems biology
- Biochemistry
Background:
- Crosslinking mass spectrometry (XMS) is a powerful technique for protein structure determination.
- Its application is expanding from structural biology to systems biology due to its sensitivity and ability to analyze complex biological samples, including whole cells.
- The effectiveness of XMS heavily relies on the design and properties of crosslinking reagents.
Purpose of the Study:
- To emphasize the need for rigorous experimental evaluation of crosslinker concepts and functional groups.
- To advocate for a data-driven approach in designing crosslinking reagents.
- To investigate the impact of reactive group kinetics and enrichment strategies on XMS.
Main Methods:
- Focus on evaluating concepts like reaction selectivity, enrichability, cleavability, and isotope-labeling in crosslinkers.
- Experimental assessment of reactive group reaction kinetics.
- Analysis of enrichment group strategies for improved XMS data.
Main Results:
- Current crosslinker design is often concept-driven rather than data-driven.
- Thorough experimental validation is crucial to understand the utility of specific functional groups and concepts.
- Reaction kinetics and enrichment strategies significantly influence XMS data quality and information density.
Conclusions:
- Crosslinker design requires a shift towards empirical data to optimize performance.
- Understanding the precise role and impact of each component in a crosslinker is essential for advancing XMS.
- Data-driven optimization of crosslinkers will enhance their utility in structural and systems biology.
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