Over the past week, I have continued to analyze the mass spectrometry (MS) data from the limited proteolysis experiment, performed on the full-length Q17 Huntingtin protein sample from Stefan Kochanek’s lab, and also assessed how this data tallies with previously published studies. To pull all of this analysis together from the past few blog posts, I have made a figure which I hope visually explains what the data shows so far. This is taken from my latest data upload (http://dx.doi.org/10.5281/zenodo.46008). You can also, as always, read about it in more detail on Zenodo.
The limited proteolysis data maps fragments which correlate well with the predicted domains of huntingtin based on InterPro analysis. It would be great to be able to confirm these domain boundaries by using a different enzyme in the digestion and this will likely be the next experiment I aim to complete.
Once I have validated these findings further, I would ideally like to repeat this experiment with a polyQ expanded full-length huntingtin protein to see if the structural organisation is changed in the disease-associated form of the protein.