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Anal Biochem. 2015 May 15;477:95-7. doi: 10.1016/j.ab.2014.11.006. Epub 2014 Nov 29.

Can your protein be sumoylated? A quick summary and important tips to study SUMO-modified proteins.

Author information

1
Department of Biology, Yeshiva University, New York, NY 10016, USA.
2
Laboratory for Macromolecular Analysis and Proteomics, Albert Einstein College of Medicine, Bronx, NY 10461, USA.
3
Department of Biology, Yeshiva University, New York, NY 10016, USA; Department of Developmental and Molecular Biology, Albert Einstein College of Medicine, Bronx, NY 10461, USA. Electronic address: vigodner@yu.edu.

Abstract

A diverse set of SUMO target proteins has been identified. Therefore, there is a growing interest in studying sumoylation and SUMO interactions in cells. When the sumoylation of a protein or a SUMO interaction is suspected, a standard co-immunoprecipitation analysis using anti-SUMO and anti-target protein antibody is usually performed as a first step. However, the identification of endogenous sumoylated proteins is challenging because of the activity of isopeptidases, and often only a small fraction of a target protein is sumoylated at a given time. Here, we briefly summarize several important steps to ensure a successful co-immunoprecipitation analysis to detect possible sumoylation.

KEYWORDS:

Co-immunoprecipitation; In vitro sumoylation; Isopeptidases; Mass spectrometry; SUMO; Sumoylation

PMID:
25454506
PMCID:
PMC4404192
DOI:
10.1016/j.ab.2014.11.006
[Indexed for MEDLINE]
Free PMC Article

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