I’m sad to report that a couple of test expressions later, we’re still no wiser as to why ACVR1 isn’t expressing. It’s worked on a couple of small scale expression test plates, but just doesn’t want to scale up anymore. We may be looking at having to remake the virus.
In the meantime, I had some old purified ACVR1A (short form R206H mutant) (a different construct) in my freezer, so I put it on the gel filtration again and set up a few plates with it the other week with some of the M4K compounds (M4K2009, M4K3007). These made some crystals (kind of ugly crystals – see below), and I had some 2 month old TGFBR1/FKBP12 plates that had some really, really fine hair like crystals on them, so I mounted all these and sent them for screening. Sadly, nothing diffracted.
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ACVR1 crystals with M4K2009
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ACVR1 with M4K2009
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ACVR1 with M4K2009
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TGFBR1 (ALK5) with FKBP12 and M4K1170
However, although not much is working in the lab at the moment, I did an outreach event on Saturday afternoon in Bonn Square in Oxford – this was Soapbox Science (https://www.iccs.org.uk/event/soapbox-science-oxford-2018 and http://soapboxscience.org/soapbox-science-2018-oxford/ and @SoapboxScienceOxf). This event aims to showcase women scientists and their work, giving us the space to engage with the public about what we do. It was a huge amount of fun. People stayed for quite a long time to listen and ask questions, and we had some truly fascinating talks.