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  • J Biol Chem. 2013-10; NULL NULL NULL
  • A soluble fragment of the tumor antigen BCL2-associated athanogene 6 (BAG-6) is essential and sufficient for inhibition of NKp30-dependent cytotoxicity of natural killer cells.

  • Binici J, Hartmann J, Herrmann J, Schreiber C, Beyer S, GÜler G, Vogel V, Tumulka F, Abele R, Mäntele W, Koch J.
  • Institute for Biomedical Research, Germany.

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... Immunoprecipitation - Tagged BAG-6686-936 proteins were mixed with IgG1-Fc fusion proteins for 1 h and incubated with 20 μl of magnetic beads covalently coated with polyhistidine tag-specific mouse monoclonal antibodies (GenScript) or StrepTactin-coated magnetic beads ...


J Biol Chem.

Abstract

Immunosurveillance of tumor cells depends on NKp30, a major activating receptor of human natural killer (NK) cells. The human BCL2-associated athanogene 6 (BAG-6, also known as BAT3; 1126 amino acids) is a cellular ligand of NKp30. To date, little is known about the molecular details of this receptor ligand system. Within the current study, we have located the binding site of NKp30 to a sequence stretch of 250 amino acids in the C-terminal region of BAG-6 (BAG-6(686-936)). BAG-6(686-936) forms a non-covalent dimer of 57-59 kDa, which is sufficient for high affinity interaction with NKp30 (KD < 100 nM). As our most important finding, BAG-6(686-936) inhibits NKp30-dependent signaling, interferon-gamma release and degranulation of NK cells in the presence of malignantly transformed target cells. Based on these data, we show for the first time that BAG-6(686-936) comprises a sub-domain of BAG-6, which is sufficient for receptor docking and inhibition of NKp30-dependent NK cell cytotoxicity as part of a tumor immune escape mechanism. These molecular insights provide an access point to restore tumor immunosurveillance by NK cells and to increase the efficacy of cellular therapies.

Keywords

BAG-6; BAT3; Immunosuppression; Innate immunity; NKp30; Natural killer (NK) cell; Tumor immunology; Tumor marker; natural killer cell receptors (NCR); tumor immune escape

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... Immunoprecipitation - Tagged BAG-6686-936 proteins were mixed with IgG1-Fc fusion proteins for 1 h and incubated with 20 μl of magnetic beads covalently coated with polyhistidine tag-specific mouse monoclonal antibodies (GenScript) or StrepTactin-coated magnetic beads ...

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