Test Expression of Huntingtin Domain Constructs in Insect Cell Culture

I am back from vacation and other distractions and now have some new and exciting data to share with everyone. Following on the previous limited proteolysis experiment where I chopped up the protein into proteolyticaly stable nuggets which allowed me to work out the potential domain boundaries of the huntingtin protein, I designed constructs which Read More …

Limited proteolysis of full-length huntingtin with chymotrypsin

I have received data from SPARC, Sick Kids Hospital Toronto, for the limited digest experiment I previously wrote about. This experiment aims to work out where the flexible linker regions of the protein structure are, therefore predicting which regions of the protein sequence correspond to folded domains.All of the experimental data is uploaded onto Zenodo. Very Read More …

Huntingtin linker sequence determination by computational methods – correspondence with Alex Holehouse

I firmly believe that the more scientists discuss their data with experts both within their field and outside of it, the better! Without peer review, constructive criticism and comprehensive evaluation of the data we all generate, we can not understand its limitations nor its value to others working in the same field. In this vein, Read More …

Domain mapping of full-length huntingtin by limited proteolysis with chymotrypsin – continued

Yesterday, I managed to overcome my eagerness to see huntingtin fragments on a gel and restained overnight with blue-silver coomassie to reveal fairly respectable bands, much more amenable to subsequent extraction from the gel and mass spectrometry analysis. All of the my write up for yesterday is on Zenodo. The team at SPARC, Sick Kids Read More …