Synthesis and testing of follow up compounds based on HAO1 fragment 6, bound at the tetramer interface.

Hello again! A few posts ago, when I shared my fragment screening results (here), I mentioned that we had one fragment that bound at the interface between two subunits of the HAO1 tetramer (figure 1 will refresh your memory). The simplest way to turn this into an inhibitor would be to make it bigger so Read More …

Purification and crystallisation of an ALK2/ACVR2/FKPB12 complex.

One problem with working with the isolated kinase domains of ALK2 and it’s type II binding partners, is that the affinity between the kinase domains alone is insufficient to allow complex formation. We know that the two proteins associate with each other in vivo and are co-localised by the domains on the outside of the Read More …

Shorter procedure to access Thieno[2,3-d]pyrimidines

Continuing with the line of delivering a chemical probe for DRAK2/STK17B kinase, we decided to revise the procedure to synthesize analogs with a thieno[2,3-d]pyrimidine core, which are intended to be used as negative control in our DRAK2 inhibition study. We shortened the synthesis of these pyrimidines to 2 steps from the starting material 6-bromo-4-chlorothieno[2,3-d]pyrimidine 1 Read More …

Screening ACVR1 inhibitors on mutant and non-mutant ACVR1 DIPG cells – effectiveness may vary

Hi there! The last month of my life was taken over by making sure my PhD first year report was beautifully polished, but I have returned with results of a small compound screen: These are all compounds that Jong Fu has already tested with his assays so we know they effectively inhibit ACVR1, but I Read More …

Validated antibodies for ALS research: the ALS-RAP project

We are part of ALS-RAP, the ALS (Amyotrophic Lateral Sclerosis) Reproducible Antibody Platform. We’re addressing one of the big conundrums to biomedical research: the lack, or the poor quality, of many antibodies used for research ranging from cell biology to histopathology. ALS, also called MND (motor neurone disease), is a group of diseases that cause Read More …

CaMKK2 Inhibitor Series

The line of up of different inhibitor series (hinge binders) for CaMKK2 that we are currently pursuing include: furopyridine, azaindole (pyrrolopyridine), quinoline, triazolopyridazine, imidazopyrazine, pyrazolopyrimidine. Figure 1. Figure 1: CaMKK2 hinge binders As part of our structure and activity relationships (SAR), we continue to build diversity on the pyridine ring for the furopyridine series, and Read More …

Introducing the team: The Gileadi Group in Oxford.

Hi! I’m Opher Gileadi, I head a group at the Structural Genomics Consortium (SGC) in Oxford. For many years, we’ve been looking at proteins involved in human. We’ve looked at a wide variety of proteins: from DNA repair enzymes and transcription factors to enzymes involved in signalling. In most cases, we focus on early work: Read More …

Update on crystal hits, XChem and more purification

Things have been really busy in the lab the last few weeks. I’ve done a couple of purifications of ACVR1 (ALK2), which I’ll post in Zenodo (getting loads of protein now), but probably won’t write up the details of every purification every single time anymore, as it’s the same process every time for this construct. Read More …

Development of a Mass Spectrometry USP5 Catalytic Activity Assay

Now that I’ve got my biophysical screening assays developed (19F NMR, SPR), as well as my hit expansion campaign started for small molecule inhibitors against the USP5 Zf-UBD, I need to be able to test if these small molecule inhibitors will antagonize the catalytic activity of USP5. It has been speculated in literature that the Read More …