Profiling Ubiquitin and Ubiquitin-like Dependent Post-translational Modifications and Identification of Significant Alterations

This protocol is an easy way to generate the profiles of ubiquitin and ubiquitin-like post-translational modifications by identifying and semi-quantifying specific identified proteins. The main advantage of this technique is the use of lentiviruses to create stable cell line of expression within a week, and the use of two tags to specifically purify proteins. In fact, this kind of post-translational modifications are involved in most biological functions, and alterations of this mechanism are found in most disease.

Therefore, identifying these alterations may serve as a prognostic and/or diagnosis and of course as molecular targets. Ubiquitin is one of the most conserved protein in eukaryotic cells and this essential for their survival. Therefore, this kind of method could be applied to any eukaryotic model, from mammals to yeast.