Identification and analysis of multiprotein complexes through chemical crosslinking
Karsten Melcher1, Hung-Ta Chen
1University of Ulster, Coleraine, United Kingdom.
Current Protocols in Cell Biology
|January 30, 2008
Summary
Chemical crosslinking helps study protein interactions within large complexes. This guide explains various crosslinking techniques and provides protocols for His6-mediated and photoinducible label transfer crosslinking for analyzing multiprotein complexes.
Area of Science:
- Biochemistry
- Molecular Biology
- Proteomics
Background:
- Chemical crosslinking is crucial for investigating protein-protein interactions in intact protein complexes.
- Analyzing large multiprotein complexes requires specialized crosslinking techniques.
Purpose of the Study:
- To guide researchers in selecting appropriate chemical crosslinking methods for specific biological questions.
- To provide practical protocols for analyzing large multiprotein complexes.
Main Methods:
- Exploration of diverse chemical crosslinking techniques with varying chemistries and scopes.
- Detailed protocols for His6-mediated crosslinking.
- Detailed protocols for photoinducible label transfer crosslinking.
Main Results:
- The study elucidates the complexities of chemical crosslinking, offering a structured approach to method selection.
- Provides practical, sample protocols for two effective crosslinking methods.
Conclusions:
- Chemical crosslinking is a powerful tool for understanding protein complex organization.
- The provided methods, His6-mediated and photoinducible label transfer crosslinking, are particularly useful for large multiprotein complex analysis.
