Making huntingtin nanobodies

Prof Ray Truant kindly shared his nanobody construct with us. This encodes a camelid derived Ig domain which binds huntingtin as he describes in this paper. This may be helpful in stabilizing our huntingtin protein samples to make it easier to work with in our structural experiments. Our cloning team recloned the nanobody DNA sequences Read More …

Post-translational modifications of insect cell derived huntingtin and success with HMGB1

Hi folks! Recently, I have been doing some work to further characterise the huntingtin protein samples I am generating in the lab. My huntingtin protein is made in insect cells and these cells process proteins they make differently to human cells. In particular, I am keen to see whether a chemical modification called phosphorylation is Read More …

Purification of high mobility group box 1 protein (HMGB1) – a huntingtin interacting protein

To generate better data for cryo-EM analysis of the huntingtin protein structure, more stable and conformationally homogeneous samples may help improve the particle averages. Purifying complexes of huntingtin with proteins which specifically bind huntingtin may promote such sample stability. High mobility group box 1 (HMGB1) is known to bind huntingtin but, to my knowledge, this interaction Read More …