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 …

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

I am continuing to try and understand the domain architecture of the huntingtin protein. In a bid to validate my previous limited proteolysis/mass spectrometry experiment, I wanted to see if I could get similar proteolytically stable fragments by digesting my remaining aliquot (courtesy of the generous Stefan Kochanek and Bin Huang) of full-length huntingtin protein Read More …

CHDI Meeting in Palm Springs – Thursday: Cell Replacement Technologies and Clinical Trial Data Analysis

The first session of the final day at the CHDI meeting focused on restoring neuronal networks. Most of these methodologies focus on the once fanciful idea that damaged neurons in the brain can be replaced with healthy new ones. Work by Ali Brivanlou using embryonic stem cells (cells which may be nurtured to grow into Read More …