Systems-wide analysis of ADP-ribosylation using quantitative mass spectrometry
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The Nielsen group develops new methodologies enabling proteome-wide characterizing of underexplored post-translational modifications. Recently the group established a methodology that allows unbiased studies of endogenous ADP-ribosylation in a systems-wide manner. Here we describe usage of the Af1521 methodology for characterization of the ADP-ribosylation regulome as a function of cellular regulation based upon stimulus, time, and enzymatic perturbations. To this end, we identified and quantified thousands of ADP-ribosylation sites and determined their temporal dynamics after stimulating human cells with various types of genotoxic stresses, and profiled the regulation of the ADP-ribosylome upon cellular deletion of HPF1 and ARH3. We furthermore demonstrate the applicability of Activated Ion Electron Transfer Dissociation (AI-ETD) for improved detection and quantification of ADP-ribosylation, hereby advancing the study of ADP-ribosylation under physiological conditions and opening up new analytical directions for the integrative analysis of the ADP-ribosylation regulome. Collectively, we present a proteomics-based characterization of the ADP-ribosylation regulome and provide new insights into the global, integrative view of cellular regulation of this important protein modification.
Date: 3 July 2020, 14:00 (Friday, 10th week, Trinity 2020)
Venue: Pathology EPA Building, off South Parks Road OX1 3UB
Venue Details: Seminar Room
Speaker: Professor Michael Nielsen (Novo Nordisk Foundation, Copenhagen)
Organising department: Sir William Dunn School of Pathology
Organiser: Dawn Gibbons (Sir William Dunn School of Pathology )
Organiser contact email address: dawn.gibbons@path.ox.ac.uk
Host: Dr Ivan Ahel (Sir William Dunn School of Pathology, University of Oxford)
Part of: Dunn School of Pathology Departmental Seminars
Booking required?: Not required
Audience: Members of the University only
Editor: Dawn Gibbons