Limited proteolysis of full-length huntingtin with chymotrypsin

I have received data from SPARC, Sick Kids Hospital Toronto, for theĀ 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

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 …

Domain mapping of full-length Q17 huntingtin – summary so far

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 sampleĀ 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 Read More …

Mass Spectrometry Data

Currently, I am continuing to work on the domain mapping experiments with huntingtin (HTT) protein. I hope that by working out more about the domain organisation of this protein, I will be able to infer information about its 3-dimensional structure (what does it look like?) and maybe its functions in the cell (what is it Read More …