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Protein interactions, post-translational modifications and topologies in human cells
Juan D Chavez1, Chad R Weisbrod, Chunxiang Zheng
1Department of Genome Science, University of Washington, Seattle, Washington 98109, USA.
Molecular & Cellular Proteomics : MCP
|January 29, 2013
Summary
Researchers used chemical cross-linking and mass spectrometry to map protein interactions in living human cells. This technique revealed 368 cross-links, offering new insights into protein structures and functions, including those with epigenetic modifications.
Area of Science:
- Biochemistry
- Molecular Biology
- Cell Biology
Background:
- Protein surface topology dictates specific biomolecular interactions essential for cellular functions.
- Current technological limitations hinder large-scale studies of protein structures and interactions within living systems.
- Chemical cross-linking offers a method for rapid stabilization of proteins and complexes, enabling in-cell structure-function relationship studies.
Purpose of the Study:
- To apply advanced chemical cross-linking and mass spectrometry techniques to analyze protein topologies and interactions in living human cells.
- To identify and characterize cross-links across various cellular compartments, including membrane, cytosolic, and nuclear proteins.
- To investigate intraprotein and interprotein cross-links in core histone proteins, particularly those with post-translational modifications.
Main Methods:
- Utilized state-of-the-art chemical cross-linking technologies.
- Employed mass spectrometry for high-throughput analysis of cross-linked proteins.
- Applied these methods to samples derived from living human cells.
Main Results:
- Identified a total of 368 cross-links from diverse cellular locations.
- Observed intraprotein and interprotein cross-links involving core histone proteins.
- Detected cross-links associated with post-translational modifications on histone proteins, relevant to epigenetic functions.
- Provided detailed structural information on known multi-protein complexes and suggested novel protein-protein interactions.
Conclusions:
- Demonstrated the effectiveness of cross-linking for direct topological measurements of post-translationally modified proteins.
- The study provides novel insights into the structural organization of proteins within living human cells.
- These findings enhance our understanding of protein complex structures and interactions, including those involved in epigenetic regulation.
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