ISSN: 0974-276X
Dorota Skowyra
Tanzania
Research Article
Broad Utility of an Affinity-enrichment Strategy for Unanchored Polyubiquitin Chains
Author(s): Jo Strachan, Barry Shaw, Varun Gopala Krishna, Daniel Scott, Juan C del Pozo, Kristine Hill, Madhav Gautam, Dorota Skowyra, Andrew D. Jacobson, Chang-Wei Liu, Neil J Oldham and Rob LayfieldJo Strachan, Barry Shaw, Varun Gopala Krishna, Daniel Scott, Juan C del Pozo, Kristine Hill, Madhav Gautam, Dorota Skowyra, Andrew D. Jacobson, Chang-Wei Liu, Neil J Oldham and Rob Layfield
Protein ubiquitination is a common post-translational modification where selected targets are covalently modified by the ubiquitin protein, often in the form of isopeptide-linked polyubiquitin chains. More recently, unanchored (i.e. non-substrate-linked) polyubiquitin chains have also been described and implicated in a range of biological processes. The development of Tandem-repeated Ubiquitin-Binding Entities (TUBEs), engineered repeats of ubiquitin-binding domains that interact non-covalently with polyubiquitin, has allowed strategies for the affinity-enrichment of ubiquitinmodified proteins to be established, in some cases with linkage specificity. Here, we demonstrate the utility of a Free Ubiquitin-Binding Entity (FUBE), based on an ubiquitin-binding domain with high specificity for the free C-terminus of ubiquitin (the Znf-UBP domain of human USP5). In contrast to TUBEs which do.. View More»
DOI:
10.4172/jpb.S7-001