Identifying new members of the huntingtin interactome with BioID and other updates

I have been busy in the lab working at the bench and training our new recruits to the Huntington’s disease research team! Welcome Jacob and Claudia!

Using a technique called BioID, we have identified proteins which are proximal or nearby to the HTT protein in cells. To date, no published literature details the use of this methodology to identify proteins which might interact with HTT so this represents a novel approach. Working in collaboration with Prof. Anne-Claude Gingras (Lunenfeld Tanenbaum Research Institute, University of Toronto) and her postdoctoral fellow Dr. Geoff Hesketh, we have used this technique to finalise a short-list of supposed HTT interacting proteins, many of which are newly described in their connection to Huntington’s disease. This is an exciting step forward to this project and I am keen to follow up all of these new leads.

Working in collaboration with Prof. Hilal Lashuel and Driss Boudeffa as well as our cryoEM collaborators Prof. Susan Lea and Dr. Justin Deme, we have analysed how altering the phosphorylation state of HTT might change the global structure. We analysed our dephosphorylated samples by cryoEM and we can see the global structure of the protein from analysis of the images collected by Susan and Justin. I am looking forward to their next update on the remaining samples.

The stringent protein purification protocol for HTT is now optimised and we used this to purify both HTT and HTT-HAP40 Q23 . These samples are ready for analysis and use in different interaction studies.

I am away from the lab for the next few weeks as I will be travelling to Europe to attend the Gordon Research Conference on Triplet Repeat Disorders in Italy. I am excited to give my talk and present my poster as well as catch up with many of my HD colleagues and collaborators.

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