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Identification of Ubiquitination-Associated Proteins Using 2D-DIGE

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Difference Gel Electrophoresis

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2596))

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Abstract

Ubiquitination is a post-translational modification, in which a small regulatory protein (~8.6 kDa) is tagged as a single moiety or as a chain to target proteins. Ubiquitination is the most versatile cellular regulatory mechanism, essential to the physiological and pathophysiological cellular events that regulate protein turnover, gene transcription, cell cycle progression, DNA repair, apoptosis, viral budding, and receptor-mediated endocytosis. Changes and abnormalities within the ubiquitination process can result in a plethora of diseases, including various cancers. The ubiquitination process is tightly controlled in a stepwise manner by four enzymes: E1 ubiquitin-activating enzymes, E2 ubiquitin-conjugating enzymes, E3 ubiquitin-ligating enzymes, and deubiquitinating proteases. Using fluorescence two-dimensional difference gel electrophoresis (2D-DIGE) to detect and quantitate cellular proteins associated with the ubiquitination process will facilitate the evaluation of this post-translational modification associated with the pathophysiological phenotype.

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Authors and Affiliations

  1. Department of Biology, Maynooth University, National University of Ireland, Maynooth, Co. Kildare, Ireland

    Paul Dowling

  2. Department of Haematology, Mater Misericordiae University Hospital, Dublin, Ireland

    Despina Bazou

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  1. Paul Dowling
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Correspondence to Despina Bazou .

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  1. Maynooth University, MAYNOOTH, Ireland

    Kay Ohlendieck

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Dowling, P., Bazou, D. (2023). Identification of Ubiquitination-Associated Proteins Using 2D-DIGE. In: Ohlendieck, K. (eds) Difference Gel Electrophoresis. Methods in Molecular Biology, vol 2596. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-2831-7_6

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  • DOI: https://doi.org/10.1007/978-1-0716-2831-7_6

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