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 has not been demonstrated in the published literature with purified recombinant protein samples – just with confocal microscopy and co-immunoprecipitation studies. To begin to investigate this interaction, I first need to express and purify HMGB1.

This experiment was not very successful! I over-expressed HMGB1 no problem but the purification failed. However, I think I know how to fix it so am planning to repeat ASAP. You can read all of the details of how I did the experiments and how I plan to improve them on Zenodo.

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