Chapter title |
Monitoring the Sensitivity of T Cell Populations Towards NAD+ Released During Cell Preparation
|
---|---|
Chapter number | 22 |
Book title |
ADP-ribosylation and NAD+ Utilizing Enzymes
|
Published in |
Methods in molecular biology, August 2018
|
DOI | 10.1007/978-1-4939-8588-3_22 |
Pubmed ID | |
Book ISBNs |
978-1-4939-8587-6, 978-1-4939-8588-3
|
Authors |
Björn Rissiek, Marco Lukowiak, Friedrich Haag, Tim Magnus, Friedrich Koch-Nolte, Rissiek, Björn, Lukowiak, Marco, Haag, Friedrich, Magnus, Tim, Koch-Nolte, Friedrich |
Abstract |
Mouse T cells express the toxin-related ecto-ADP-ribosyltransferase ARTC2 that catalyzes the posttranslational ADP-ribosylation of cell surface proteins by transferring the ADP-ribose group of its substrate nicotinamide adenine dinucleotide (NAD+) to arginine residues of its target proteins. One well known target of ARTC2 is the ATP-gated P2X7 ion channel. ADP-ribosylation of P2X7 induces gating of the channel, calcium influx, ecto-domain shedding, phosphatidylserine externalization, and finally cell death. Previous studies have shown that the ARTC2 substrate NAD+ is released during T cell preparation. Since P2X7 is differentially expressed among T cell subpopulations, preparation-related ADP-ribosylation has a strong impact on the vitality of T cells that express high levels of P2X7. With this chapter we provide a protocol to monitor the consequences of preparation-related P2X7 ADP-ribosylation on T cells using regulatory T cells as generic T cell subpopulation known to express high levels of P2X7. However, this protocol could be easily adapted to other T cell populations. |
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