PIAS1 (full length) Purification

Huntington’s Disease (HD) is a hereditary neurodegenerative disease. The cause of this disease is a CAG repeat extension in the HTT Gene. This extension is translated into an elongation of exon 1 in Huntingtin protein. Although we know the cause, the mechanism of this disease still eludes us. A part of the difficulty is our lack of understanding of the role of normal HTT in our cells. We at the SGC have set out to explore the HTT interactome to further our understanding of the role of HTT. This will give us a better basis for further study of mutant Huntingtin. Our first step on this journey was to conduct a literature review alongside a BioID experiment to come up with a list of putative huntingtin interaction partners. One of the hits on the list was PIAS1. This means that we must obtain a pure sample of PIAS1 for use in future experiments to further validate the claim that these proteins interact. Previously, I purified a shorter construct of PIAS1 in E. coli : 10.5281/zenodo.3367282. At the SGC we have access to many different constructs of the same protein. The previously purified product was only a portion of the full protein. For physiological relevance, it is important to test full length proteins so we decided to order another purification of PIAS1, and this time try to purify the whole protein.Funding Acknowledgment: The SGC is a registered charity (number 1097737) that receives funds from AbbVie, Bayer Pharma AG, Boehringer Ingelheim, Canada Foundation for Innovation, Eshelman Institute for Innovation, Genome Canada through Ontario Genomics Institute [OGI-055], Innovative Medicines Initiative (EU/EFPIA) [ULTRA-DD grant no. 115766], Janssen, Merck KGaA, Darmstadt, Germany, MSD, Novartis Pharma AG,Innovation and Science (MRIS), Pfizer, São Paulo Research Foundation-FAPESP, Takeda, and Wellcome