Published March 20, 2019 | Version v1
Dataset Open

Preparation of dephosphorylated HTT samples for cryoEM analysis 2019/03/20

Creators

  • 1. Structural Genomics Consortium, University of Toronto
  • 2. Princess Margaret Cancer Centre and Department of Medical Biophysics, University of Toronto, Toronto, Ontario M5G 1L7, Canada

Description

Project: High resolution structural analysis of purified HTT samples

Experiment: Preparation of dephosphorylated HTT samples for cryoEM analysis

Date completed:­ 2019/03/20

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. 

Notes

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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Preparation of dephosphorylated HTT samples for cryoEM analysis 20190320.pdf

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