Post-translational modification of proteins offers a rapid route to change the activity of crucial factors within the cell. One of the more drastic post-translational modifications, in terms of effect on biochemical properties, is the covalent attachment of the small protein ubiquitin, to a target factor. The labile nature of some post-translational modifications puts obstacles in the path of attempting to detect modified species of most proteins. Indeed, ubiquitination can be rapidly reversed by the action of a large family of DUBs (deubiquitinating enzymes), most of which are cysteine proteases. This, taken together with the rapid proteasomal degradation of some species of ubiquitinated proteins, results in difficulties in detecting modified targets. In this review, practical approaches developed for the detection, purification and characterization of ubiquitinated proteins are reviewed. After a brief appraisal of the use of histidine-tagged ubiquitin, focus is placed on development of UBD (ubiquitin-binding domain)–ubiquitin affinity purification.
Conference Article| September 19 2008
Efficient approaches for characterizing ubiquitinated proteins
Manuel S. Rodríguez
Manuel S. Rodríguez 1
1Ubiquitin-Like Molecules and Cancer Laboratory, Proteomics Unit, CIC bioGUNE (Asociacion Centro de Investigación Cooperativa en Biociencias), CIBERehd, Bizkaia Technology Park, Building 801A, 48160 Derio, Spain
1To whom correspondence should be addressed (email email@example.com).
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Roland Hjerpe, Manuel S. Rodríguez; Efficient approaches for characterizing ubiquitinated proteins. Biochem Soc Trans 1 October 2008; 36 (5): 823–827. doi: https://doi.org/10.1042/BST0360823
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