1. Identification of the Mouse T Cell ADP-Ribosylome Uncovers ARTC2.2 Mediated Regulation of CD73 by ADP-Ribosylation
- Author
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Mario Leutert, Yinghui Duan, Riekje Winzer, Stephan Menzel, Eva Tolosa, Tim Magnus, Michael O. Hottiger, Friedrich Koch-Nolte, Björn Rissiek, University of Zurich, Leutert, Mario, and Rissiek, Björn
- Subjects
Adenosine monophosphate ,T cell ,Immunology ,T cells ,Nicotinamide adenine dinucleotide ,CD8-Positive T-Lymphocytes ,chemistry.chemical_compound ,Mice ,ADP-Ribosylation ,medicine ,Cytotoxic T cell ,Animals ,Immunology and Allergy ,IL-2 receptor ,5'-Nucleotidase ,ARTC2.2 ,Original Research ,ADP Ribose Transferases ,Mice, Knockout ,2403 Immunology ,Chemistry ,HEK 293 cells ,RC581-607 ,NAD ,10226 Department of Molecular Mechanisms of Disease ,Cell biology ,medicine.anatomical_structure ,Membrane protein ,CD73 ,2723 Immunology and Allergy ,570 Life sciences ,biology ,NAD+ kinase ,Immunologic diseases. Allergy - Abstract
Mouse T cells express the ecto-ADP-ribosyltransferase ARTC2.2, which can transfer the ADP-ribose group of extracellular nicotinamide adenine dinucleotide (NAD+) to arginine residues of various cell surface proteins thereby influencing their function. Several targets of ARTC2.2, such as P2X7, CD8a and CD25 have been identified, however a comprehensive mouse T cell surface ADP-ribosylome analysis is currently missing. Using the Af1521 macrodomain-based enrichment of ADP-ribosylated peptides and mass spectrometry, we identified 93 ADP-ribsoylated peptides corresponding to 67 distinct T cell proteins, including known targets such as CD8a and CD25 but also previously unknown targets such as CD73. We evaluated the impact of ADP-ribosylation on the capability of CD73 to generate adenosine from adenosine monophosphate. Our results show that extracellular NAD+reduces the enzymatic activity of CD73 HEK cells co-transfected with CD73/ARTC2.2. Importantly, NAD+significantly reduced CD73 activity on WT CD8 T cells compared to ARTC2ko CD8 T cells or WT CD8 T cells treated with an ARTC2.2-blocking nanobody. Our study provides a comprehensive list of T cell membrane proteins that serve as targets for ADP-ribosylation by ARTC2.2 and whose function may be therefore affected by ADP-ribosylation.
- Published
- 2021
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