title: Nucleotide-Induced Membrane-Proximal Proteolysis Controls the Substrate Specificity of T Cell Ecto-ADP-Ribosyltransferase ARTC2.2.
abstract: ARTC2.2 is a toxin-related, GPI-anchored ADP-ribosyltransferase expressed by murine T cells. In response to NAD(+) released from damaged cells during inflammation, ARTC2.2 ADP-ribosylates and thereby gates the P2X7 ion channel. This induces ectodomain shedding of metalloprotease-sensitive cell surface proteins. In this study, we show that ARTC2.2 itself is a target for P2X7-triggered ectodomain shedding. We identify the metalloprotease cleavage site 3 aa upstream of the predicted GPI anchor attachment site of ARTC2.2. Intravenous injection of NAD(+) increased the level of enzymatically active ARTC2.2 in serum, indicating that this mechanism is operative also under inflammatory conditions in vivo. Radio-ADP-ribosylation assays reveal that shedding refocuses the target specificity of ARTC2.2 from membrane proteins to secretory proteins. Our results uncover nucleotide-induced membrane-proximal proteolysis as a regulatory mechanism to control the substrate specificity of ARTC2.2.
authors.affiliation: Institute of Immunology, University Medical Center Hamburg-Eppendorf, D20246 Hamburg, Germany;
authors.forename: Stephan
authors.initials: S
authors.lastname: Menzel
authors.affiliation: Institute of Immunology, University Medical Center Hamburg-Eppendorf, D20246 Hamburg, Germany; Department of Neurology, University Medical Center Hamburg-Eppendorf, D20246 Hamburg, Germany;
authors.forename: Björn
authors.initials: B
authors.lastname: Rissiek
authors.affiliation: Institute of Immunology, University Medical Center Hamburg-Eppendorf, D20246 Hamburg, Germany; Department of Diagnostic Radiology, University Medical Center Hamburg-Eppendorf, D20246 Hamburg, Germany; and.
authors.forename: Peter
authors.initials: P
authors.lastname: Bannas
authors.affiliation: Institute of Experimental Medicine, Systematic Proteome Research Group, Christian-Albrechts-Universität, D24105 Kiel, Germany.
authors.forename: Thomas
authors.initials: T
authors.lastname: Jakoby
authors.affiliation: Institute of Immunology, University Medical Center Hamburg-Eppendorf, D20246 Hamburg, Germany;
authors.forename: Maria
authors.initials: M
authors.lastname: Miksiewicz
authors.affiliation: Institute of Immunology, University Medical Center Hamburg-Eppendorf, D20246 Hamburg, Germany;
authors.forename: Nicole
authors.initials: N
authors.lastname: Schwarz
authors.affiliation: Institute of Immunology, University Medical Center Hamburg-Eppendorf, D20246 Hamburg, Germany;
authors.forename: Marion
authors.initials: M
authors.lastname: Nissen
authors.affiliation: Institute of Immunology, University Medical Center Hamburg-Eppendorf, D20246 Hamburg, Germany;
authors.forename: Friedrich
authors.initials: F
authors.lastname: Haag
authors.affiliation: Institute of Experimental Medicine, Systematic Proteome Research Group, Christian-Albrechts-Universität, D24105 Kiel, Germany.
authors.forename: Andreas
authors.initials: A
authors.lastname: Tholey
authors.affiliation: Institute of Immunology, University Medical Center Hamburg-Eppendorf, D20246 Hamburg, Germany; nolte@uke.de.
authors.forename: Friedrich
authors.initials: F
authors.lastname: Koch-Nolte
mesh_terms: D051722:ADAM Proteins / Q000378:metabolism; D000072198:ADAM17 Protein; D036002:ADP Ribose Transferases / Q000097:blood / Q000235:genetics / Q000378:metabolism*; D000246:Adenosine Diphosphate Ribose / Q000378:metabolism; D000818:Animals; D045744:Cell Line, Tumor; D002462:Cell Membrane / Q000187:drug effects / Q000378:metabolism; D005434:Flow Cytometry; D017261:Glycosylphosphatidylinositols / Q000378:metabolism; D057809:HEK293 Cells; D006801:Humans; D019041:L-Selectin / Q000235:genetics / Q000378:metabolism; D008565:Membrane Proteins / Q000235:genetics / Q000378:metabolism*; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D008822:Mice, Transgenic; D009243:NAD / Q000378:metabolism* / Q000494:pharmacology; D059748:Proteolysis / Q000187:drug effects; D058486:Receptors, Purinergic P2X7 / Q000235:genetics / Q000378:metabolism; D013379:Substrate Specificity; D016176:T-Lymphocyte Subsets / Q000201:enzymology / Q000378:metabolism; D013601:T-Lymphocytes / Q000201:enzymology* / Q000378:metabolism
chemical_list: D017261:Glycosylphosphatidylinositols; D008565:Membrane Proteins; D058486:Receptors, Purinergic P2X7; D009243:NAD; D019041:L-Selectin; D000246:Adenosine Diphosphate Ribose; D036002:ADP Ribose Transferases; C000603470:ARTC2.2 protein, mouse; C449339:Art2b protein, mouse; D051722:ADAM Proteins; D000072198:ADAM17 Protein