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Further down the line I want to be able to test the activation of the BMP signalling pathway downstream of the ALK2 receptor. For this I plan to use the alkaline phosphatase gene as a readout – it’s a well known gene activated downstream of BMP signalling so a lot of easy to use assays Read More …
Hello everyone and happy New Year! I really, really wanted to end the year on a high note in December before I went off on holiday, but as it turned out, I didn’t have enough time to set up 18 plates and write a blog post on the last day. So I’ll just have to Read More …
In an effort to develop clinical compounds for Diffused Intrinsic Pontine Glioma (DIPG) treatment, new analogues of ACVR1/ALK2 inhibitors are continuously synthesised by Ontario Institute for Cancer Research (OICR) and Charles River Laboratories (CRL). I will provide prompt feedback of the cellular assay results to guide their design of new compounds. I determined the potency Read More …
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 …
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 …
So I reported here a couple of weeks ago that I’d obtained datasets for ACVR1 bound to M4K2117 and M4K2121, at 1.25 and 2 Å resolution, respectively, and I put up a link to the data. I’m now refining the data and got some pretty good structures so far. These are what the current stats look Read More …
So, the week has been super busy, but all for good reasons this time!! I’m absolutely thrilled to say that amongst all the salt crystals I found in my crystal plates, I also found the crystals below, one of which diffracted to 1.25 Å. This is definitely the highest resolution I’ve ever worked with. It’s amazing! Read More …
As mentioned in my last post, I got 26 mg of ACVR1 on my last purification (as shown in my Zenodo log here). This is fabulous as it has allowed me to follow up a number of conditions for crystallisation. In particular, one of these hits was in G10 of my composite screen. I’ve therefore Read More …
Now that our workhorse ACVR1 construct is expressing again, I’m getting loads of protein – 26 mg on the last 3 L scale up! Because of this, I’ve been really busy in the lab trying to get things back up to speed again. While waiting for the ACVR1 to work, I purified some TGFBR1 (ALK5) Read More …
Structure Solution for BMPR1B/FKBP12 complex bound to M4K2009. Good news everybody! From the crystals I sent to the Diamond Light Source (a synchrotron, or particle accelerator, that gives very high quality x-rays) last week, I got a data set for BMPR1B/FKBP12 with M4K2009 bound to it! This is one of the compounds that the M4K Read More …
