Structure Solution for BMPR1B/FKBP12 complex bound to M4K2009.

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 …

Complex formation of FKBP12/BMPR1A and FKBP12/BMPR1B and subsequent crystallisation.

Having previously purified BMPR1A (Alk3) and BMPR1B (Alk6), I spent some time purifying FKBP12 to form complexes with each of the receptors. FKBP12 binds to BMPR1A and BMPR1B via a similar GS domain to that seen in ACVR1, on the N-terminal side of the kinase domain and may help stabilize the structures and thus help Read More …

Purification of ACVR2, Alk3 (BMPR1A) and Alk6 (BMPR1B)

I had a couple of weeks off over August but I was back last week ready to crack on with work. As you’ve probably been following the ongoing saga, Ros and I have been struggling with expression of our key constructs. She had some good news on that on her latest post which has some Read More …

Purification and crystallisation of ALK2 (R206H mutation) with M4K Pharma compounds.

As part of an ongoing collaboration with Meds4Kids Pharma (M4KPharma), we are testing a range of compounds identified and generated by them as being of interest in developing a molecule specific against ALK2 that has the right properties help treat some cases of DIPG. This also has significant implications for development of a drug that Read More …

Complex formation of XIAP with ACVR2.

  It’s been a very busy few weeks, a fair amount of lab work but also a fair bit of outreach work (which is the other part of my job) and so I’ve sadly neglected to write my poor blog post. However,  now I’ve got the chance to write a bit about what I’ve been Read More …

Pull down of XIAP with type I & II receptors. Tm shifts of M4K Pharma compounds with Alk2, Alk5 and RIPK2.

XIAP and its interaction with type I and type II receptors. Having spent time over the past few weeks purifying all the components needed for a pull down with XIAP, I did the experiment that looked for interactions between XIAP and various type I and type II receptors in both the un-phosphorylated and phosphorylated form. Read More …

Protein purification and Tm shift experiments.

Purification of proteins for a pull down experiment between XIAP and various type I and type II receptors. XIAP is known to bind to the kinase RIPK2. RIPK2 is one of the few kinase proteins that shows overlap of reactivity to some of the compounds that we’ve been looking at in the context of ALK2 Read More …

Structure deposition of ALK2 structures and structural analysis of ALK2/FKBP12 complex with PK010710 and PK012055

Deposition: In the past two weeks my work has mostly been full of dealing with structures. The first thing I needed to do was deposit some structures of ALK2 co-crystallised with three drug molecules that I had solved for our collaborators Swen Hoelder and Liam Hudson at ICR, who had been developing compounds against ALK2. Read More …

Crystal mounting of ALK2 with FKBP12 and six compounds with subsequent data collection

One of the more unpredictable parts of crystallography is also one of the most fundamental – will the protein crystallise? Despite decades of work done on protein crystallisation, and numerous ‘tricks of the trade’ to maximise the chances, whether a protein will actually form a crystal is still relatively random.  With ALK2 we’ve crystallised it Read More …

Co-Crystallisation of the ALK2/FKBP12 complex with compounds from M4K and others.

New compounds developed by Meds4Kids (M4K) and other collaborators against ALK2 would benefit from being crystallised so we can look at the structure of the complex and explain exactly how these compounds bind. This helps us work out how to improve them for both potency and selectivity and explain why some bind better than others. Read More …