For the past few days, I have been working on optimizing a fluorescence polarization (FP) displacement assay for USP5 Zf-UBD. This assay will be used to test if small molecules can displace a fluorescence-labelled ubiquitin peptide from USP5. I started with screening different assay conditions with a small set of buffers. The method and data Read More …

Before I can begin characterizing the USP5 protein, I first needed to make some protein sample to work with. I expressed and purified the zinc-finger domain protein in bacteria and ended up with a high yield of a sample which is very pure. All of my methods and data from this experiment can be found Read More …

Ubiquitination can mark proteins for degradation, alter their cellular location, affect their activity, and promote or prevent protein interactions. Ubiquitin specific proteases (USPs) are the largest family of deubiquitinase enzymes. USP5 disassembles unanchored ubiquitin chains. This prevents the accumulation of ubiquitin chains which would otherwise compete against ubiquitinated proteins for degradation as well as replenishes the Read More …