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Analysis of dephosphorylated HTT samples for cryoEM analysis and further sample preparation 2019/04/15

Harding, Rachel; Deme, Justin; Lea, Susan; Arrowsmith, Cheryl; Edwards, Aled

Project: High resolution structural analysis of purified HTT samples

Experiment: Analysis of dephosphorylated HTT samples for cryoEM analysis and further sample preparation

Date completed:­ 2019/04/15

Rationale: We now have a good understanding of the global structure of HTT when in complex with HAP40 and our MALS and SAXS data suggest that apo HTT is likely self-associating and heterogenous in nature, despite high levels of purity. Dephosphorylating HTT from Sf9 cell production could alter its global structure sufficiently to allow high resolution structure determination by cryoEM. 

Dr. Harding is the recipient of the Huntington's Disease Society of America Berman Topper Career Development Fellowship which funds and supports this research, in addition to generous funding from the CHDI Foundation and the Huntington Society of Canada. 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, Ontario Ministry of Research, Innovation and Science (MRIS), Pfizer, São Paulo Research Foundation-FAPESP, Takeda, and Wellcome.
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