Fine definition of the epitopes on the human gp91 phox /NOX2 for the monoclonal antibodies CL-5 and 48.

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  • Additional Information
    • Source:
      Publisher: Elsevier Country of Publication: Netherlands NLM ID: 1305440 Publication Model: Print-Electronic Cited Medium: Internet ISSN: 1872-7905 (Electronic) Linking ISSN: 00221759 NLM ISO Abbreviation: J Immunol Methods Subsets: MEDLINE
    • Publication Information:
      Publication: Amsterdam : Elsevier
      Original Publication: Amsterdam, North-Holand,
    • Subject Terms:
      Epitope Mapping* ; Epitopes*; Antibodies, Monoclonal/*immunology ; NADPH Oxidase 2/*immunology; Animals ; Antibody Specificity ; CHO Cells ; COS Cells ; Chlorocebus aethiops ; Cricetulus ; Glycosylation ; HL-60 Cells ; Humans ; Mice ; NADPH Oxidase 2/genetics ; NADPH Oxidase 2/metabolism ; Protein Processing, Post-Translational ; RAW 264.7 Cells ; Receptors, Immunologic/genetics ; Receptors, Immunologic/immunology ; Receptors, Immunologic/metabolism ; Superoxides/metabolism
    • Abstract:
      Superoxide-producing NADPH oxidase, gp91 phox /NOX2, in phagocytes plays a critical role in the host defenses against pathogens. Moreover, gp91 phox /NOX2 contributes to the oxidative stress in endothelial cells. Therefore, investigating the level of gp91 phox /NOX2 with immunoblotting is important for estimating the amount of superoxide produced. Here, we showed that the epitopes in human gp91 phox /NOX2 recognized by monoclonal antibodies (mAbs) CL-5 and 48 were in amino acids 132-147 and 136-144, respectively. Although the epitopes were close to the N-glycosylation sites, N-glycan maturation did not affect mAbs recognition. When Pro-136 was substituted with Arg, the corresponding mouse residue, human gp91 phox /NOX2 was not recognized by mAbs CL-5 and 48; however, the substitution did not affect gp91 phox /NOX2-based oxidase activity. This finding explains why these mAbs specifically recognize the human but not mouse gp91 phox /NOX2. Hence, these mAbs are useful for investigating the level of gp91 phox /NOX2 without amino acid substitutions in the epitopes.
      (Copyright © 2021 Elsevier B.V. All rights reserved.)
    • Contributed Indexing:
      Keywords: Chronic granulomatous disease (CGD); Endothelial cell; Monoclonal antibody; Phagocyte; gp91(phox)/NOX2
    • Accession Number:
      0 (Antibodies, Monoclonal)
      0 (Epitopes)
      0 (Pirb protein, mouse)
      0 (Receptors, Immunologic)
      11062-77-4 (Superoxides)
      EC 1.6.3.- (CYBB protein, human)
      EC 1.6.3.- (NADPH Oxidase 2)
    • Publication Date:
      Date Created: 20211231 Date Completed: 20220131 Latest Revision: 20220131
    • Publication Date:
      20240829
    • Accession Number:
      10.1016/j.jim.2021.113213
    • Accession Number:
      34971634