Preliminary experiments to assess the suitability of DLS for protein interaction studies with huntingtin-HAP40 2019/08/12

Harding, Rachel; Arrowsmith, Cheryl; Edwards, Aled

doi:10.5281/zenodo.3383304

Published August 12, 2019 | Version v1

Dataset Open

Preliminary experiments to assess the suitability of DLS for protein interaction studies with huntingtin-HAP40 2019/08/12

1. Structural Genomics Consortium, University of Toronto

Project: Biophysical investigation of the HTT interactome

Experiment: Preliminary experiments to assess the suitability of DLS for protein interaction studies with huntingtin-HAP40

Date completed: 2019/08/12

Rationale: HTT is a multi-valent protein interactor and scaffold. It is likely that many of the proteins HTT interacts with form weak or transient interactions. One method which may be fruitful to detect this type of interaction is dynamic light scattering – a method which allows in solution calculation of protein particle radius. HTT-HAP40 Q23 (https://zenodo.org/record/3383265), PACSIN1 (https://zenodo.org/record/3256589) and PFN1A (kindly donated by Opher Gileadi and Tracy Keates) were analysed in a dilution series by DLS to assess signal to noise and concentration dependency of protein radius and polydispersity.

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. Thanks to Opher Gileadi and Tracy Keates for providing the PFN1 protein for this experiment. Thanks also to Greg Wasney, Max Jorgensen and Claudia Alvarez for help with DLS experiments. Thanks to Matt Lee and Liz Doherty for experimental ideas and strategy planning.

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