Chamberlain Lab

Slide 1

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The S-acylation field is currently hindered by a lack of information about which DHHC enzymes are active against which S-acylated substrates, and how enzyme-substrate specificity is encoded. To investigate DHHC-substrate interaction specificity, along with using standard approaches, such as immunoprecipitations and pull-downs, we have recently implemented the yeast-based protein interaction platform Split-Ubiquitin System (SUS)

 

SUS is a variation of the classic Yeast Two-Hybrid System and is used for assessment of interaction between a membrane protein having a cytoplasmic C-terminus (bait) and any other protein (prey). As the name suggests, it relies on ubiquitin being split into two, with one half fused in-between the bait and a transcriptional activator, and the other half appended to the prey. Interaction of bait and prey will lead to the assembly of whole ubiquitin, subsequent release of the transcriptional activator (by ubiquitin-specific proteases), and transcription of reporter genes essential for yeast growth on selective growth medium.

 

SUS01