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Bimolecular fluorescence complementation to assay the interactions of ubiquitylation enzymes in living yeast cells

Methods in Molecular Biology, ISSN: 1064-3745, Vol: 1449, Page: 223-241
2016
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Book Chapter Description

Ubiquitylation is a versatile posttranslational protein modification catalyzed through the concerted action of ubiquitin-conjugating enzymes (E2s) and ubiquitin ligases (E3s). These enzymes form transient complexes with each other and their modification substrates and determine the nature of the ubiquitin signals attached to their substrates. One challenge in the field of protein ubiquitylation is thus to identify the E2-E3 pairs that function in the cell. In this chapter, we describe the use of bimolecular fluorescence complementation to assay E2-E3 interactions in living cells, using budding yeast as a model organism.

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