A growing team of groundbreaking scientists around the world are now sharing their lab notebooks online
Today’s post describes testing if ZnF-UBD ligands are able to antagonize USP5 catalytic activity using a ubiquitin-rhodamine110 (UbRho110) assay, which was developed and optimized by Leanna Smith of the Biophysics group at SGC Toronto. You can see details on Zenodo. The UbRho110 assay is a fluorescence intensity assay that monitors the enzymatic activity of deubiquitinating Read More …
In a previous post, I described the expression and purification of full length (FL) USP5 constructs and also determined that zinc finger ubiquitin-binding (ZnF-UBD) ligands are able to displace ubiquitin in both the ZnF-UBD and the FL protein in a fluorescence displacement assay. In today’s post, I describe using a surface plasmon resonance (SPR) assay Read More …
Five months ago, I made my first post outlining the goals of my project here at the SGC. I am writing again today to update you on the progress I have made toward those goals. As a brief overview, I was working on finding Huntingtin (HTT) interacting proteins, but why would this be important? If Read More …
I’ve been working recently on purifying some proteins for use in future experiments and wanted to explain a little bit about what I need each construct for as they’re all for different things. Purification, as always, is a time consuming business but vital for any biophysical experiment that I do given all my work is Read More …
Man, it has gotten cold and windy, its only been a month, and already we have gone from +25 degree sunshine, to overcast, sub-10 degree temperatures, with rain, true Irish weather. The joys of Fall as winter approaches. I hope everyone is doing well, I enjoyed my short hiatus from the notebook, but am now Read More …
As part of SGC-UNC’s scaffold hopping strategy to identify new series of compounds active against CaMKK2, we recently submitted a select number of compounds differing in hinge-binder for CaMKK2 enzyme inhibition assay. The hinge-binders included thienopyridine, furopyridine, quinoline, thienopyrimidines, pyrimidine, N-methyl azaindoles and quinazoline, Figure 1. Others are azaindoles, triazolopyridazine, pyrazolopyridine, imidazopyridine etc. Figure 1: Read More …
Recombinant protein expression is a prominent molecular technique for structural biology studies. For expressing recombinant proteins, a host organism is utilized as protein production factories. To obtain the desired recombinant proteins, we first design a recombinant DNA, which involves the information of protein to be produced. Then, we clone the desired DNA fragments into a Read More …
Hello folks, another day another deposition. That makes it sound horribly commonplace but actually Ros put in immense effort into crystallising and solving these proteins and it’s my job just to add the final polish (fix those last stubborn geometry errors), give it a once over to make sure there’s nothing obviously wrong, and whip Read More …
