Method for the Purification of Endogenous Unanchored Polyubiquitin Chains.

Autor:	Scott D; School of Life Sciences, Queen's Medical Centre, University of Nottingham, Nottingham, NG7 2UH, UK., Strachan J; Institute of Cell Biology, University of Edinburgh, Edinburgh, UK., Krishna VG; School of Life Sciences, Queen's Medical Centre, University of Nottingham, Nottingham, NG7 2UH, UK., Shaw B; School of Life Sciences, Queen's Medical Centre, University of Nottingham, Nottingham, NG7 2UH, UK., Tooth DJ; School of Life Sciences, Queen's Medical Centre, University of Nottingham, Nottingham, NG7 2UH, UK., Searle MS; School of Chemistry, University of Nottingham, University Park, Nottingham, NG7 2RD, UK.; Centre for Biomolecular Sciences, University of Nottingham, University Park, Nottingham, NG7 2RD, UK., Oldham NJ; School of Chemistry, University of Nottingham, University Park, Nottingham, NG7 2RD, UK., Layfield R; School of Life Sciences, Queen's Medical Centre, University of Nottingham, Nottingham, NG7 2UH, UK. robert.layfield@nottingham.ac.uk.
Jazyk:	angličtina
Zdroj:	Methods in molecular biology (Clifton, N.J.) [Methods Mol Biol] 2016; Vol. 1449, pp. 203-13.
DOI:	10.1007/978-1-4939-3756-1_11
Abstrakt:	Unanchored polyubiquitin chains are endogenous non-substrate linked ubiquitin polymers which have emerging roles in the control of cellular physiology. We describe an affinity purification method based on an isolated ubiquitin-binding domain, the ZnF_UBP domain of the deubiquitinating enzyme USP5, which permits the selective purification of mixtures of endogenous unanchored polyubiquitin chains that are amenable to downstream molecular analyses. Further, we present methods for detection of unanchored polyubiquitin chains in purified fractions.
Databáze:	MEDLINE
Externí odkaz:	Zobrazit plný text záznamu