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Multidimensional protein identification technology (MudPIT) analysis of ubiquitinated proteins in plants
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Maor, R., Jones, Alexandra M., Nühse, Thomas S., Studholme, David J., Peck, Scott C. and Shirasu, Ken (2007) Multidimensional protein identification technology (MudPIT) analysis of ubiquitinated proteins in plants. Molecular & Cellular Proteomics, Volume 6 (Number 4). pp. 601-610. doi:10.1074/mcp.M600408-MCP200 ISSN 1535-9476.
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Official URL: http://dx.doi.org/10.1074/mcp.M600408-MCP200
Abstract
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.
Item Type: | Journal Article | ||||
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Divisions: | Faculty of Science, Engineering and Medicine > Science > Life Sciences (2010- ) | ||||
Journal or Publication Title: | Molecular & Cellular Proteomics | ||||
Publisher: | American Society for Biochemistry and Molecular Biology | ||||
ISSN: | 1535-9476 | ||||
Official Date: | 1 April 2007 | ||||
Dates: |
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Volume: | Volume 6 | ||||
Number: | Number 4 | ||||
Page Range: | pp. 601-610 | ||||
DOI: | 10.1074/mcp.M600408-MCP200 | ||||
Status: | Peer Reviewed | ||||
Publication Status: | Published | ||||
Access rights to Published version: | Restricted or Subscription Access |
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