title: The Stalk Domain of NKp30 Contributes to Ligand Binding and Signaling of a Preassembled NKp30-CD3ζ Complex.
abstract: The natural cytotoxicity receptor (NCR) NKp30 (CD337) is a key player for NK cell immunosurveillance of infections and cancer. The molecular details of ligand recognition and its connection to CD3ζ signaling remain unsolved. Here, we show that the stalk domain (129KEHPQLGAGTVLLLR143) of NKp30 is very sensitive to sequence alterations, as mutations lead to impaired ligand binding and/or signaling capacity. Surprisingly, the stalk domains of NKp30 and NKp46, another NCR employing CD3ζ for signaling, were not exchangeable without drastic deficiencies in folding, plasma membrane targeting, and/or ligand-induced receptor signaling. Further mutational studies, N-glycosylation mapping, and plasma membrane targeting studies in the absence and presence of CD3ζ suggest two interconvertible types of NCR-CD3ζ assemblies: 1) a signaling incompetent structural NKp30-CD3ζ complex and 2) a ligand-induced signaling competent NKp30-CD3ζ complex. Moreover, we propose that ligand binding triggers translocation of Arg-143 from the membrane interface into the membrane to enable alignment with oppositely charged aspartate residues within CD3ζ and activation of CD3ζ-signaling.
authors.affiliation: From the Institute of Medical Microbiology and Hygiene, University of Mainz Medical Center, 55131 Mainz, Germany.|the Institute for Tumor Biology and Experimental Therapy, Georg-Speyer-Haus, 60596 Frankfurt am Main, Germany.
authors.forename: Stefanie
authors.initials: S
authors.lastname: Memmer
authors.affiliation: From the Institute of Medical Microbiology and Hygiene, University of Mainz Medical Center, 55131 Mainz, Germany.|the Institute for Tumor Biology and Experimental Therapy, Georg-Speyer-Haus, 60596 Frankfurt am Main, Germany.
authors.forename: Sandra
authors.initials: S
authors.lastname: Weil
authors.affiliation: the Institute for Tumor Biology and Experimental Therapy, Georg-Speyer-Haus, 60596 Frankfurt am Main, Germany.
authors.forename: Steffen
authors.initials: S
authors.lastname: Beyer
authors.affiliation: the Institute for Molecular Medicine, Goethe-University Frankfurt am Main, 60528 Frankfurt am Main, Germany.
authors.forename: Tobias
authors.initials: T
authors.lastname: Zöller
authors.affiliation: the Institute for Tumor Biology and Experimental Therapy, Georg-Speyer-Haus, 60596 Frankfurt am Main, Germany.|the Institute for Microbiology, ETH Zürich, 8093 Zürich, Switzerland.
authors.forename: Eike
authors.initials: E
authors.lastname: Peters
authors.affiliation: the Division of Molecular Biotechnology and Gene Therapy, Paul-Ehrlich-Institut, 63225 Langen, Germany, and.
authors.forename: Jessica
authors.initials: J
authors.lastname: Hartmann
authors.affiliation: the Institute for Molecular Medicine, Goethe-University Frankfurt am Main, 60528 Frankfurt am Main, Germany.
authors.forename: Alexander
authors.initials: A
authors.lastname: Steinle
authors.affiliation: From the Institute of Medical Microbiology and Hygiene, University of Mainz Medical Center, 55131 Mainz, Germany, joachim.koch@uni-mainz.de.|the Institute for Tumor Biology and Experimental Therapy, Georg-Speyer-Haus, 60596 Frankfurt am Main, Germany.|the LOEWE Excellence Center for Cell and Gene Therapy, 60590 Frankfurt am Main, Germany.
authors.forename: Joachim
authors.initials: J
authors.lastname: Koch
mesh_terms: D000818:Animals; D017252:CD3 Complex / Q000235:genetics / Q000378:metabolism*; D002462:Cell Membrane / Q000235:genetics / Q000378:metabolism*; D006367:HeLa Cells; D006801:Humans; D051379:Mice; D055627:Natural Cytotoxicity Triggering Receptor 1 / Q000235:genetics / Q000378:metabolism; D055628:Natural Cytotoxicity Triggering Receptor 3 / Q000235:genetics / Q000378:metabolism*; D000072417:Protein Domains; D015398:Signal Transduction / Q000502:physiology*
chemical_list: D017252:CD3 Complex; C115335:CD3E protein, human; C112260:NCR1 protein, human; C431710:NCR3 protein, human; D055627:Natural Cytotoxicity Triggering Receptor 1; D055628:Natural Cytotoxicity Triggering Receptor 3
keywords: cell signaling; cell surface receptor; membrane protein; natural killer cells (NK cells); protein-protein interaction; receptor modification; receptor regulation