Examining post-translational modification-mediated protein-protein interactions using a chemical proteomics approach
Xiang Li1, Emily A Foley, Shigehiro A Kawashima
1Laboratory of Chemistry and Cell Biology, New York, New York 10065, USA.
Protein Science : a Publication of the Protein Society
|January 3, 2013
Summary
We expanded CLASPI, a chemical proteomics method, to identify protein interactions regulated by post-translational modifications (PTMs) like methylation and phosphorylation in yeast and human cells.
Area of Science:
- Chemical proteomics
- Molecular and Cellular Biology
- Post-translational modifications
Background:
- Post-translational modifications (PTMs) regulate cellular processes by controlling protein-protein interactions.
- Identifying PTM-dependent interactions is challenging due to their dynamic and often weak nature.
Purpose of the Study:
- To extend the CLASPI (cross-linking-assisted and stable isotope labeling in cell culture-based protein identification) approach for profiling PTM-dependent protein-protein interactions.
- To demonstrate CLASPI's applicability in model organisms and for different PTMs.
Main Methods:
- Adaptation of CLASPI for analyzing methylation-dependent interactions in fission yeast, focusing on trimethylated histone H3 lysine-9 (H3K9Me₃).
- Application of CLASPI to study phosphorylation-dependent interactions, specifically identifying proteins binding to phosphorylated histone H3 threonine-3 (H3T3-Phos).
- Investigation of the interplay between H3T3-Phos and trimethylated histone H3 lysine-4 (H3K4Me₃) using the CLASPI approach.
Main Results:
- CLASPI successfully profiled H3K9Me₃-dependent interactions in fission yeast.
- The method identified known (survivin) and novel proteins (MCAK, KIF2A) interacting with H3T3-Phos.
- CLASPI revealed insights into the crosstalk between H3T3-Phos and H3K4Me₃.
Conclusions:
- The CLASPI approach is a versatile chemical proteomics tool for studying PTM-mediated protein-protein interactions.
- CLASPI can be broadly applied across different organisms and PTMs, including methylation, phosphorylation, and their crosstalk.
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