ACVR1 Crystals taken to the Diamond Light Source (Beamline I04.) – no data sets collected.

Following up on the crystals from my last post I sent them along on the last Diamond trip we had to beamline I04 but regrettably didn’t get any usable datasets. Perhaps fortunately there is a possible explanation for this that isn’t just ‘the crystals were a bit too rubbish’. We send our crystals to the Diamond Read More …

Re-Purification of ALK2 for crystallisation with different compounds from M4K.

I’ve been lucky enough to have a student (Mira) in the lab working with me for the past few weeks and so I’d like to write about the work that we’ve been doing together. We spent some time cleaning up an old sample of ALK2 that Ros had left over in the freezer to set Read More …

The Benefits of Conference attendance: BMP Signalling in Cancer II.

I’ve got a few things to catch up with on the blog – one of them is to talk about a conference I attended in Oxford recently. I attended the Biochemical Society BMP Signalling in Cancer II meeting which was held in St. Annes College, Oxford. At this meeting I presented a poster on the Read More …

Phosphorylation over time and phosphomapping experiments to investigate the influence of ALK2 mutations on activity.

In this post I’d like to return to looking at my work on phosphorylation. Phosphorylation is a key signalling method for activating the BMP pathway and inducing activity. Specifically activation of ALK2 by phosphorylation leads to phosphorylation of SMAD1 which leads to gene transcription that ultimately is responsible for bone formation. In this post I’m Read More …

ZFYVE9C purification and association test with TGFBR1 and TGFBR2

ZFYVE9C (Otherwise known as SARA) is a scaffold protein involved in the association of various protein components as part of the TGFB signalling pathway. The equivalent in the BMP signalling pathway is a protein known as Endofin so studying SARA (which is slightly easier) might give us useful insights for both proteins. For more details Read More …

A summary of my ALK2 project goals.

I thought I’d spend a blog post just talking about the various paths of my research and try and tie it all together. A lot of my experiments do dot around these various arms of investigation and so one experimental post doesn’t necessarily tie in directly with the previous one or the next one and 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 …

Generating mutations in the GS domain of ALK2

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 …

Phosphorylation of SMAD1 – rates as a function of Alk2 mutations and type II association.

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 …

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 …