Kei Miyano
   Department   Kawasaki Medical School  Kawasaki Medical School, Department of Natural Sciences,
   Position   Assistant Professor
Article types 原著
Language English
Peer review Peer reviewed
Title Fine definition of the epitopes on the human gp91phox/NOX2 for the monoclonal antibodies CL-5 and 48
Journal Formal name:Journal of Immunological Methods
Abbreviation:J Immunol Methods
ISSN code:00221759
Domestic / ForeginForegin
Volume, Issue, Page 501
Author and coauthor Kawai C, Miyano K, Okamoto S, Yamauchi A, Kuribayashi F.
Publication date 2022/02
Summary Superoxide-producing NADPH oxidase, gp91phox/NOX2, in phagocytes plays a critical role in the host defenses against pathogens. Moreover, gp91phox/NOX2 contributes to the oxidative stress in endothelial cells. Therefore, investigating the level of gp91phox/NOX2 with immunoblotting is important for estimating the amount of superoxide produced. Here, we showed that the epitopes in human gp91phox/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 gp91phox/NOX2 was not recognized by mAbs CL-5 and 48; however, the substitution did not affect gp91phox/NOX2-based oxidase activity. This finding explains why these mAbs specifically recognize the human but not mouse gp91phox/NOX2. Hence, these mAbs are useful for investigating the level of gp91phox/NOX2 without amino acid substitutions in the epitopes.
PMID 34971634