Analysis of Ubc13 and Uev1A Interaction and Standardization of Expression and Purification protocols for Uba1, TRAF6, HLTF and Ubiquitin

Jain, Diksha (2017) Analysis of Ubc13 and Uev1A Interaction and Standardization of Expression and Purification protocols for Uba1, TRAF6, HLTF and Ubiquitin. Masters thesis, Indian Institute of Technology Hyderabad.

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Abstract

Ubiquitin is a central regulator of a cell ’ s functions. These functions include DNA repair, protein kinase activation, and chromatin dynamics. Ubiquitin forms several kinds of polyubiquitin chains, which signals to recruit proteins harboring ubiquitin- binding domains, and ubiquitin receptors to execute specific biological functions. Interleukin-1 pathway for protein kinase activation, which requires Lys- 63 polybiquitin chains is studied here. It requires Ubc13 and Uev1A as Ubiquitin conjugating enzyme, and TRAF6 as Ubiquitin Ligase Enzyme. Ubc13-Uev1A interaction is analyzed using pulldown assays. For Lys-63 polyubiquitination, Uba1, ubiquitin activating enzyme is used as E1. Th e expression and purification protocols for all these proteins E1(Uba1), E3(TRAF6, HLTF), and ubiquitin are standardized in this study.

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IITH Creators:
IITH CreatorsORCiD
Item Type: Thesis (Masters)
Uncontrolled Keywords: TD892, HLFT, TRAF6, Ubiquitin
Subjects: Others > Biotechnology
Divisions: Department of Biotechnology
Depositing User: Team Library
Date Deposited: 12 Jul 2017 11:41
Last Modified: 12 Jul 2017 11:41
URI: http://raiith.iith.ac.in/id/eprint/3372
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