Mittal, Anshumali and Shakya, Binita (2018) Synthesis and purification of linkage-specific polyubiquitin chains of distinct length for structural studies. In: ANALYTICAL BIOCHEMISTRY, 559 . pp. 1-4.
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Abstract
Polyubiquitylation is one of the most versatile post-translational modifications involved in the regulation of numerous intracellular signaling processes. An assembly procedure that is simple, robust, and efficient to synthesize and purify linkage-specific polyubiquitin chains of defined length at a preparative scale is required in biophysical and structural studies. Here, we have optimized known enzymatic procedures in the form of a protocol to obtain multi-milligrams of Lys48-and Lys63-linked polyubiquitin chain types with more than 99% purity. Mass spectrometry (ESI/MS) analysis of K48- and K63-linked diubiquitin confirmed that the enzymes used in the preparation generated homogeneous linkages with no promiscuity.
Item Type: | Journal Article |
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Publication: | ANALYTICAL BIOCHEMISTRY |
Publisher: | ACADEMIC PRESS INC ELSEVIER SCIENCE |
Additional Information: | Copy right for this article belong to ACADEMIC PRESS INC ELSEVIER SCIENCE |
Keywords: | Ubiqutin; K48-and K63 linked polyubiquitin chains; Chromatography |
Department/Centre: | Division of Biological Sciences > Molecular Biophysics Unit |
Date Deposited: | 15 Oct 2018 15:24 |
Last Modified: | 15 Oct 2018 15:24 |
URI: | http://eprints.iisc.ac.in/id/eprint/60877 |
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