Nucleotide-Induced Membrane-Proximal Proteolysis Controls the Substrate Specificity of T Cell Ecto-ADP-Ribosyltransferase ARTC2.2

Stephan Menzel; Björn Rissiek; Peter Bannas; Thomas Jakoby; Maria Miksiewicz; Nicole Schwarz; Marion Nissen; Friedrich Haag; Andreas Tholey; Friedrich Koch-Nolte

doi:10.4049/jimmunol.1401677

Nucleotide-Induced Membrane-Proximal Proteolysis Controls the Substrate Specificity of T Cell Ecto-ADP-Ribosyltransferase ARTC2.2

J Immunol. 2015 Sep 1;195(5):2057-66. doi: 10.4049/jimmunol.1401677. Epub 2015 Jul 24.

Authors

Affiliations

¹ Institute of Immunology, University Medical Center Hamburg-Eppendorf, D20246 Hamburg, Germany;
² Institute of Immunology, University Medical Center Hamburg-Eppendorf, D20246 Hamburg, Germany; Department of Neurology, University Medical Center Hamburg-Eppendorf, D20246 Hamburg, Germany;
³ Institute of Immunology, University Medical Center Hamburg-Eppendorf, D20246 Hamburg, Germany; Department of Diagnostic Radiology, University Medical Center Hamburg-Eppendorf, D20246 Hamburg, Germany; and.
⁴ Institute of Experimental Medicine, Systematic Proteome Research Group, Christian-Albrechts-Universität, D24105 Kiel, Germany.
⁵ Institute of Immunology, University Medical Center Hamburg-Eppendorf, D20246 Hamburg, Germany; nolte@uke.de.

PMID: 26209623
DOI: 10.4049/jimmunol.1401677

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.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

ADAM Proteins / metabolism
ADAM17 Protein
ADP Ribose Transferases / blood
ADP Ribose Transferases / genetics
ADP Ribose Transferases / metabolism*
Adenosine Diphosphate Ribose / metabolism
Animals
Cell Line, Tumor
Cell Membrane / drug effects
Cell Membrane / metabolism
Flow Cytometry
Glycosylphosphatidylinositols / metabolism
HEK293 Cells
Humans
L-Selectin / genetics
L-Selectin / metabolism
Membrane Proteins / genetics
Membrane Proteins / metabolism*
Mice, Inbred C57BL
Mice, Knockout
Mice, Transgenic
NAD / metabolism*
NAD / pharmacology
Proteolysis / drug effects
Receptors, Purinergic P2X7 / genetics
Receptors, Purinergic P2X7 / metabolism
Substrate Specificity
T-Lymphocyte Subsets / enzymology
T-Lymphocyte Subsets / metabolism
T-Lymphocytes / enzymology*
T-Lymphocytes / metabolism

Substances

Glycosylphosphatidylinositols
Membrane Proteins
Receptors, Purinergic P2X7
NAD
L-Selectin
Adenosine Diphosphate Ribose
ADP Ribose Transferases
ARTC2.2 protein, mouse
Art2b protein, mouse
ADAM Proteins
ADAM17 Protein