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 …

Cloning, expression and purification of brand new huntingtin constructs

Nearly all experiments to date have used full-length huntingtin cloned by Ji-Joon Song and Ihn Sir Seong, who kindly shared their expression constructs with us. Since then, we have cloned our own full-length expression constructs which can express full-length huntingtin protein in both insect cells AND mammalian cells. The wild-type (Q23) N-terminally tagged vector generated Read More …

Investigating huntingtin DNA binding

Studies which look for genome wide-associated risk factors (GWAS) have a shown a link between Huntington’s disease and DNA repair proteins as well as other DNA related proteins including splicing and transcription factors. Huntingtin was previously described as a DNA binding protein but I have not found studies using purified huntingtin protein and DNA samples in Read More …