Shaunak Raval1, Vlad Sarpe2, Terry Zhang3, Rosa Viner3, Albert Konijnenberg4, Andreas Huhmer3, David C. Schriemer1,2
Purpose: - Study the conformational changes accompanying the in-vitro activation of 20S proteasome catalytic activity in the presence of low concentration of SDS
Methods: HDX-MS was used to study the perturbation of conformational dynamics of Rabbit 20S proteasome complex in the presence of sodium dodecyl sulfate (SDS). The complete data analysis including peptide identification and determination of deuterium uptake was performed using HDX-MS package in the Mass Spec Studio (v2.0).
Results: Newly implemented peptide identification and filtering workflow in MS Studio (v2.0) allows for one step enrichment of HDX ready peptide features for downstream analysis. We utilized this functionality in the analysis of conformational dynamics of 20S proteasome in the presence of SDS. Our results indicate that the low concentration of SDS has significant impact on the structure of proteasome with possibility of partial unfolding of the subunits.
1Department of Chemistry, 2Department of Biochemistry and Molecular Biology, University of Calgary, Calgary, Alberta, Canada Thermo Fisher Scientific, 3San Jose, US, 4Eindhoven, The Netherlands