|
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 / Foregin | Foregin |
| 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. |
| DOI | https://doi.org/10.1016/j.jim.2021.113213 |
| PMID | 34971634 |