title: Functional analysis of ligand-binding and signal transduction domains of CD69 and CD23 C-type lectin leukocyte receptors.
abstract: CD69 and CD23 are leukocyte receptors with distinctive pattern of cell expression and functional features that belong to different C-type lectin receptor subfamilies. To assess the functional equivalence of different domains of these structurally related proteins, a series of CD69/CD23 chimeras exchanging the carbohydrate recognition domain, the neck region, and the transmembrane and cytoplasmic domains were generated. Biochemical analysis revealed the importance of the neck region (Cys68) in the dimerization of CD69. Functional analysis of these chimeras in RBL-2H3 mast cells and Jurkat T cell lines showed the interchangeability of structural domains of both proteins regarding Ca2+ fluxes, serotonin release, and TNF-alpha synthesis. The type of the signal transduced mainly relied on the cytoplasmic domain and was independent of receptor oligomerization. The cytoplasmic domain of CD69 transduced a Ca2+-mediated signaling that was dependent on the extracellular uptake of Ca2+. Furthermore, a significant production of TNF-alpha was induced through the cytoplasmic domain of CD69 in RBL-2H3 cells, which was additive to that promoted via FcepsilonRI, thus suggesting a role for CD69 in the late phase of reactions mediated by mast cells. Our results provide new important data on the functional equivalence of homologous domains of these two leukocyte receptors.
authors.affiliation: Servicio de Inmunología, Hospital de la Princesa, Universidad Autónoma de Madrid, Diego de León, Spain.
authors.forename: D
authors.initials: D
authors.lastname: Sancho
authors.forename: A G
authors.initials: AG
authors.lastname: Santis
authors.forename: J L
authors.initials: JL
authors.lastname: Alonso-Lebrero
authors.forename: F
authors.initials: F
authors.lastname: Viedma
authors.forename: R
authors.initials: R
authors.lastname: Tejedor
authors.forename: F
authors.initials: F
authors.lastname: Sánchez-Madrid
mesh_terms: D000818:Animals; D015703:Antigens, CD / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D000945:Antigens, Differentiation, T-Lymphocyte / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D020013:Calcium Signaling / Q000235:genetics / Q000276:immunology; D016207:Cytokines / Q000096:biosynthesis; D003593:Cytoplasm / Q000235:genetics / Q000276:immunology; D006801:Humans; D007249:Inflammation / Q000276:immunology; D019169:Jurkat Cells; D037102:Lectins / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D037181:Lectins, C-Type; D008024:Ligands; D008407:Mast Cells / Q000276:immunology / Q000378:metabolism; D011485:Protein Binding / Q000235:genetics / Q000276:immunology; D017434:Protein Structure, Tertiary / Q000235:genetics; D051381:Rats; D017455:Receptors, IgE / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D011975:Receptors, Mitogen / Q000276:immunology / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000138:chemical synthesis / Q000378:metabolism; D015398:Signal Transduction / Q000235:genetics / Q000276:immunology*; D014407:Tumor Cells, Cultured; D000099192:CD69 Antigens
chemical_list: D015703:Antigens, CD; D000945:Antigens, Differentiation, T-Lymphocyte; D016207:Cytokines; D037102:Lectins; D037181:Lectins, C-Type; D008024:Ligands; D017455:Receptors, IgE; D011975:Receptors, Mitogen; D011993:Recombinant Fusion Proteins; D000099192:CD69 Antigens