Dual NADPH oxidases DUOX1 and DUOX2 synthesize NAADP and are necessary for Ca2+ signaling during T cell activation

  • Sci Signal. 2021 Nov 16;14(709):eabe3800. doi: 10.1126/scisignal.abe3800.
Feng Gu  1 ,  Aileen Krüger  1 ,  Hannes G Roggenkamp  1 ,  Rick Alpers  1 ,  Dmitri Lodygin  2 ,  Vincent Jaquet  3 ,  Franziska Möckl  1 ,  Lola C Hernandez C  1 ,  Kai Winterberg  1 ,  Andreas Bauche  1 ,  Anette Rosche  1 ,  Helmut Grasberger  4 ,  John Y Kao  4 ,  Daniel Schetelig  5 ,  René Werner  5 ,  Katrin Schröder  6 ,  Michael Carty  7 ,  Andrew G Bowie  7 ,  Samuel Huber  8 ,  Chris Meier  9 ,  Hans-Willi Mittrücker  10 ,  Joerg Heeren  1 ,  Karl-Heinz Krause  3 ,  Alexander Flügel  2 ,  Björn-Philipp Diercks  1 ,  Andreas H Guse  1
Affiliations
  • 1. Calcium Signaling Group, Department of Biochemistry and Molecular Cell Biology, University Medical Center Hamburg-Eppendorf, 20246 Hamburg, Germany.
  • 2. Institute for Neuroimmunology and Multiple Sclerosis Research, University Medical Center Göttingen, 37075 Göttingen, Germany.
  • 3. Department of Pathology and Immunology, University of Geneva, 1211 Geneva 4, Switzerland.
  • 4. Department of Internal Medicine, Division of Gastroenterology, University of Michigan, Ann Arbor, MI 48109, USA.
  • 5. Department of Computational Neuroscience, University Medical Center Hamburg-Eppendorf, Hamburg 20246, Germany.
  • 6. Institute of Cardiovascular Physiology, Goethe-Universität, 60590 Frankfurt, Germany.
  • 7. School of Biochemistry and Immunology, Trinity Biomedical Sciences Institute, Trinity College Dublin, Dublin 2, Ireland.
  • 8. Department of Gastroenterology with Sections Infectiology and Tropical Medicine, University Medical Center Hamburg-Eppendorf, 20246 Hamburg, Germany.
  • 9. Organic Chemistry, University of Hamburg, 20146 Hamburg, Germany.
  • 10. Department of Immunology, University Medical Center Hamburg-Eppendorf, 20246 Hamburg, Germany.
PMID: 34784249 DOI: 10.1126/scisignal.abe3800
Abstract

The formation of CA2+ microdomains during T cell activation is initiated by the production of nicotinic acid Adenine dinucleotide phosphate (NAADP) from its reduced form NAADPH. The reverse reaction—NAADP to NAADPH—is catalyzed by glucose 6-phosphate dehydrogenase (G6PD). Here, we identified NADPH oxidases NOX and DUOX as NAADP-forming Enzymes that convert NAADPH to NAADP under physiological conditions in vitro. T cells express NOX1, NOX2, and, to a minor extent, DUOX1 and DUOX2. Local and global CA2+ signaling were decreased in mouse T cells with double knockout of Duoxa1 and Duoxa2 but not with knockout of NOX1 or NOX2. CA2+ microdomains in the first 15 s upon T cell activation were significantly decreased in Duox2−/− but not in DUOX1−/− T cells, whereas both DUOX1 and DUOX2 were required for global CA2+ signaling between 4 and 12 min after stimulation. Our findings suggest that a DUOX2- and G6PD-catalyzed redox cycle rapidly produces and degrades NAADP through NAADPH as an inactive intermediate.