Hi folks! This past month or so I have been busy getting a manuscript draft prepared. This article details the HTT constructs we have made and shared with the community, how they can be used to make pure huntingtin protein of different polyQ lengths and characterisation of the protein samples that we made.
We think this is important because:
- Freely available tools and reagents to make HTT are critical for the HD research community to do important biochemical studies to try and further understand this protein
- Having HD tools with appropriate polyQ lengths is really important and the entry clones we have made and shared allow anyone to easily make even more HTT constructs of whatever expansion they like
- Our HTT constructs allow scalable production of HTT in multiple eukaryotic expression systems so you can choose to make protein in mammalian or insect cells and scale the production to make as much or as little as you need
- We have completed some biophysical analysis of the HTT protein with and without its binding partner HAP40, gleaning further insight into some of the characteristics of these samples
Fitting SAXS data to structural models of HTT-HAP40 complex - understanding the full-length HTT molecule
A) Experimental SAXS profiles (black circles) plotted with the theoretical profile for the EM structure of the HTT-HAP40 complex (red line) and the predicted profile averaged over the optimal Sparse Ensemble Selection (SES) ensemble (blue line). B) Surface representation of the EM structure as well as C) the most populated (~44 %) solution model and D) the complete ensemble of the HTT-HAP40 complex are shown with surface representation of the globular regions and backbone trace of the flexible regions (colour key as in panel B).
Want to know more?
- You can find all of the data contributing to the manuscript on Zenodo.
- The manuscript draft, or preprint, is on bioRxiv.
- AND if you want to make HTT samples yourself, the constructs can be found on Addgene.
Feedback and comments are thoroughly welcome!