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In Vitro SUMOylation Assay to Study SUMO E3 Ligase Activity
Published on: January 29, 2018
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Can your protein be sumoylated? A quick summary and important tips to study SUMO-modified proteins
Yuxuan Xiao1, Daniel Pollack1, Edward Nieves2
1Department of Biology, Yeshiva University, New York, NY 10016, USA.
Analytical Biochemistry
|December 3, 2014
Summary
Detecting protein sumoylation via co-immunoprecipitation is challenging due to enzyme activity. This guide outlines key steps for successful analysis of sumoylated proteins and SUMO interactions in cells.
Area of Science:
- Cellular biology
- Molecular mechanisms of protein modification
Background:
- Growing interest in studying protein sumoylation and SUMO interactions.
- Standard co-immunoprecipitation is the initial step for detecting sumoylation.
- Identifying endogenous sumoylated proteins is difficult due to isopeptidase activity and low modification levels.
Purpose of the Study:
- To summarize essential steps for successful co-immunoprecipitation.
- To aid in the detection of possible protein sumoylation.
Main Methods:
- Co-immunoprecipitation assays.
- Use of anti-SUMO and anti-target protein antibodies.
Main Results:
- Challenges in identifying endogenous sumoylated proteins are highlighted.
- Key procedural steps for effective co-immunoprecipitation are presented.
Conclusions:
- Proper methodology is crucial for reliable detection of sumoylation.
- The summarized steps aim to improve the success rate of co-immunoprecipitation experiments for studying sumoylated proteins.
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