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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 …
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 …
This week one of the things I’ve been working on is the final set of mutations in the GS domain of Alk2 to help us unpick the influence of phosphorylation on function. That, however, is a bit of a mouthful so let’s unpack it a bit. I said in my last blog post that we 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 …
One of the questions to be asked when looking at the mutation in ALK2 that causes FOP, is how exactly does that mutation cause the disease. ALK2 we know is part of the BMP signalling pathway responsible for bone formation and the mutation results in excess bone formation – thus we can summarise that somehow 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 …
