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Synthesis and purification of linkage-specific polyubiquitin chains of distinct length for structural studies

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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Official URL: http://dx.doi.org/10.1016/j.ab.2018.08.007

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
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
Depositing User: Id for Latest eprints
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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