Engineering and structural characterization of a linear polyubiquitin-specific antibody - PubMed (original) (raw)
. 2012 May 4;418(3-4):134-44.
doi: 10.1016/j.jmb.2011.12.053. Epub 2011 Dec 29.
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- PMID: 22227388
- DOI: 10.1016/j.jmb.2011.12.053
Free article
Engineering and structural characterization of a linear polyubiquitin-specific antibody
Marissa L Matsumoto et al. J Mol Biol. 2012.
Free article
Abstract
Polyubiquitination is an essential posttranslational modification that plays critical roles in cellular signaling. PolyUb (polyubiquitin) chains are formed by linking the carboxyl-terminus of one Ub (ubiquitin) subunit to either a lysine residue or the amino-terminus of an adjacent Ub. Linkage through the amino-terminus results in linear polyubiquitination that has recently been demonstrated to be a key step in nuclear factor κB activation; however, tools to study linear chains have been lacking. We therefore engineered a linear-linkage-specific antibody that is functional in Western blot, immunoprecipitation, and immunofluorescence applications. A crystal structure of the linear-linkage-specific antibody Fab fragment in complex with linear diubiquitin provides molecular insight into the nature of linear chain specificity. We use the antibody to demonstrate that linear polyUb is up-regulated upon tumor necrosis factor α stimulation of cells, consistent with a critical role in nuclear factor κB signaling. This antibody provides an essential tool for further investigation of the function of linear chains.
Copyright © 2011. Published by Elsevier Ltd.
Comment in
- New tools for ubiquitin signaling.
Keusekotten K, Komander D. Keusekotten K, et al. J Mol Biol. 2012 May 4;418(3-4):129-30. doi: 10.1016/j.jmb.2012.02.032. Epub 2012 Feb 27. J Mol Biol. 2012. PMID: 22381406 No abstract available.
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