Experiment Details

Exp 103

Publication Information

Maor et al., 2007

PubMed ID: 17272265

No external accession available

Abstract

Mol Cell Proteomics. 2007 Apr;6(4):601-10. doi: 10.1074/mcp.M600408-MCP200. Epub 
2007 Jan 31.

Multidimensional protein identification technology (MudPIT) analysis of 
ubiquitinated proteins in plants.

Maor R(1), Jones A, Nühse TS, Studholme DJ, Peck SC, Shirasu K.

Author information:
(1)The Sainsbury Laboratory, John Innes Centre, Colney Lane, Norwich NR4 7UH, 
United Kingdom.

Protein conjugation with ubiquitin, known as ubiquitination, is a key regulatory 
mechanism to control protein abundance, localization, and activity in eukaryotic 
cells. To identify ubiquitin-dependent regulatory steps in plants, we developed 
a robust affinity purification/identification system for ubiquitinated proteins. 
Using GST-tagged ubiquitin binding domains, we performed a large scale affinity 
purification of ubiquitinated proteins from Arabidopsis cell suspension culture. 
High molecular weight ubiquitinated proteins were separated by SDS-PAGE, and the 
trypsin-digested samples were then analyzed by a multidimensional protein 
identification technology (MudPIT) system. A total of 294 proteins specifically 
bound by the GST-tagged ubiquitin binding domains were identified. From these we 
determined 85 ubiquitinated lysine residues in 56 proteins, confirming the 
enrichment of the target class of proteins. Our data provide the first view of 
the ubiquitinated proteome in plants. We also provide evidence that this 
technique can be broadly applied to the study of protein ubiquitination in 
diverse plant species.

DOI: 10.1074/mcp.M600408-MCP200
PMID: 17272265 [Indexed for MEDLINE]