Related Experiment Video
Updated: Jun 10, 2026
10:01
Combining Chemical Cross-linking and Mass Spectrometry of Intact Protein Complexes to Study the Architecture of Multi-subunit Protein Assemblies
Published on: November 28, 2017
Crosslinking combined with mass spectrometry for structural proteomics
Evgeniy V Petrotchenko1, Christoph H Borchers
1University of Victoria Proteomics Centre, 3101-4464 Markham Street, Victoria, British Columbia, Canada V8Z7X8.
Mass Spectrometry Reviews
|August 24, 2010
Summary
Crosslinking combined with mass spectrometry provides detailed protein structures. Advances in reagents and software are overcoming challenges, making this technique more accessible for structural biology research.
Area of Science:
- Biochemistry
- Structural Biology
- Analytical Chemistry
Background:
- Crosslinking combined with mass spectrometry (CX-MS) is an emerging technology for elucidating protein and protein complex structures.
- Widespread adoption of CX-MS has been hindered by inherent technical challenges.
- Recent advancements in mass spectrometry instrumentation, novel crosslinking reagents, and specialized data processing software are addressing these limitations.
Purpose of the Study:
- To review recent literature on specialized crosslinking reagents and approaches for mass spectrometry-based applications.
- To assess critical features of crosslinking reagents for optimal mass spectrometric performance.
- To summarize design requirements for crosslinking reagents suitable for mass spectrometric detection and analysis.
Main Methods:
- Review of recent scientific literature focusing on crosslinking reagents and mass spectrometry.
- Assessment of reagent features including isotopic coding, cleavability, affinity groups, linker structure, and reactive groups.
- Analysis of design considerations for mass spectrometry-compatible crosslinking agents.
Main Results:
- Improvements in mass spectrometry instrumentation and software are enhancing CX-MS capabilities.
- Development of new crosslinking reagents with tailored features is crucial for improved performance.
- Key reagent characteristics such as isotopic labeling and specific reactive groups are vital for effective mass spectrometric analysis.
Conclusions:
- CX-MS is becoming a powerful tool for protein structure determination.
- Continued development of specialized crosslinking reagents is essential for advancing the field.
- Optimized reagent design is critical for maximizing the utility of CX-MS in structural biology.
Related Concept Videos
01:21Tandem Mass Spectrometry
Tandem mass spectrometry is a technique that uses multiple mass analyzers in series to obtain a higher selectivity and reduce chemical noise during analyte detection. Instruments with multiple analyzers separated by an interaction cell enable secondary fragmentation and selected study of the fragment ions.Secondary fragmentations occur in the interaction cell and can be induced by various factors. Fragmentation induced by collision with inert gases, such as N2, Ar, He, etc., is called...
01:33Peptide Identification Using Tandem Mass Spectrometry
Tandem mass spectrometry, also known as MS/MS or MS2, is an analytical technique that employs two mass analyzers. Essentially it is a series of mass spectrometers that helps isolate a particular biomolecule and then helps study its chemical properties.
This technique helps gather information regarding the protein from which the peptide was obtained and to study the peptides’ amino acid sequence. Identifying peptides from a complex mixture is an important component of the growing field of...
This technique helps gather information regarding the protein from which the peptide was obtained and to study the peptides’ amino acid sequence. Identifying peptides from a complex mixture is an important component of the growing field of...
01:21Mass Spectrometry: Complex Analysis
Mass spectrometry is an important technique for the identification of pure compounds. However, it has some limitations for the analysis of complex mixtures, often due to excessive fragmentation making the spectrum too complicated to decipher. Mass spectrometry can be combined with suitable separation methods in sequence, forming hyphenated methods, which are useful in the analysis of complex mixtures.
GC–MS is a powerful hyphenated method commonly used in forensics and environmental...
GC–MS is a powerful hyphenated method commonly used in forensics and environmental...
01:33Proteomics
A proteome is the entire set of proteins that a cell type produces. We can study proteomes using the knowledge of genomes because genes code for mRNAs, and the mRNAs encode proteins. Although mRNA analysis is a step in the right direction, not all mRNAs are translated into proteins.
Proteomics is the study of proteomes' function. It involves the large-scale systematic study of the proteome to denote the protein complement expressed by a genome. Scientist Mark Wilkins coined the term proteomics...
Proteomics is the study of proteomes' function. It involves the large-scale systematic study of the proteome to denote the protein complement expressed by a genome. Scientist Mark Wilkins coined the term proteomics...