Multidimensional Protein Identification Technology (MudPIT) Analysis of Ubiquitinated Proteins in Plants

• DOI: 10.1074/mcp.m600408-mcp200
• Publication Date: 2007-02-03T08:09:26Z
• Authors: Rudy Maor; Alex Jones; Thomas S. Nühse; David J. Studholme; Scott C. Peck; Ken Shirasu
• Keywords: Identification

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 steps plants, we developed robust affinity purification/identification system for ubiquitinated proteins. Using GST-tagged ubiquitin binding domains, performed large scale purification of proteins from Arabidopsis cell suspension culture. High molecular weight were separated by SDS-PAGE, the trypsin-digested samples then analyzed multidimensional identification technology (MudPIT) system. A total 294 specifically bound domains identified. From these determined 85 lysine residues 56 proteins, confirming enrichment target class Our data provide first view proteome plants. We also evidence that this technique can be broadly applied study ubiquitination diverse plant species.