This past week I have been trying to pin down some of the problems I have been having with making huntingtin proteins. Whilst the full-length protein prep is optimised, making smaller regions of the protein has been somewhat problematic with inconsistent yields between preps. This past week with the help of our excellent eukaryotic expression system team (Alma, Ashley and Brittany), we have managed to successfully purify large (milligram) quantities of the huntingtin protein spanning amino acids 80-428. The sample is fairly pure so I hope to do some follow up work on it this week. The write up can be read on Zenodo.
Our collaborators in Oxford, Prof Susan Lea and Dr Justin Deme, have been to Basel to collect a large-scale dataset for our huntingtin full-length Q23 cryo grids on the Krios instrument at the Novartis site with Dr Christian Wiesmann and Dr Sandra Jacob. It is great to have the help and support of the Novartis electron microscopy team on this open notebook science project. I can’t wait to see how the data look once we’ve transferred it back and have started processing.
In the interim, here is a screen shot of some particles (an actual photo taken on Susan’s phone!):
More to come soon….