RRC ID 52655
著者 Kawai C, Yamauchi A, Kuribayashi F.
タイトル Monoclonal antibody 7D5 recognizes the R147 epitope on the gp91phox , phagocyte flavocytochrome b558 large subunit.
ジャーナル Microbiol Immunol
Abstract Human phagocyte flavocytochrome b558 (Cyt b), the catalytic center of nicotinamide adenine dinucleotide phosphate oxidase, consists of a heavily glycosylated large subunit (gp91phox ; Nox2) and a small subunit (p22phox ). Cyt b is a membrane-spanning complex enzyme. Chronic granulomatous disease (CGD) is predominantly caused by a mutation in the CYBB gene encoding gp91phox on the X-chromosome. Because the phagocytes of patients with CGD are not able to generate the superoxide anion, these patients are susceptible to severe infections that can be fatal. It has been suggested that the extracellular region of gp91phox is necessary for and critical to forming the epitope of mAb 7D5 and that 7D5 provides a useful tool for rapid screening of X-linked CGD by FACS. To further elucidate the mAb 7D5 epitope on human gp91phox , chimeric DNA expressed human and mouse gp91phox recombinant protein were constructed. The fusion proteins were immunostained for mAb 7D5 and analyzed by FACS and western blot analysis. The 143 ELGDRQNES151 region was found to reside at the extracellular surface on human gp91phox and to be an important epitope for the interaction with mAb 7D5, as analyzed by FACS analysis. In particular, amino acid R147 is a unique epitope on the membrane-associated Cyt b for mAb 7D5. In conclusion, it is proposed that FACS analysis using mAb 7D5 is a valuable tool for early diagnosis of CGD.
巻・号 62(4)
ページ 269-280
公開日 2018-4-1
DOI 10.1111/1348-0421.12584
PMID 29573449
MeSH Amino Acid Sequence Animals Antibodies, Monoclonal / chemistry Antibodies, Monoclonal / immunology* Cell Line Cytochrome b Group / immunology* Epitopes / analysis Epitopes / chemistry Epitopes / immunology* Granulomatous Disease, Chronic / immunology HL-60 Cells Humans Membrane Glycoproteins / chemistry Membrane Glycoproteins / genetics Mice Mutation NADPH Oxidase 2 / biosynthesis NADPH Oxidase 2 / chemistry NADPH Oxidase 2 / genetics NADPH Oxidase 2 / immunology* NADPH Oxidases / immunology* Phagocytes / immunology* Protein Domains RAW 264.7 Cells Recombinant Fusion Proteins / genetics Sequence Analysis, Protein Sequence Homology, Amino Acid Superoxides / metabolism
IF 1.566
引用数 4
リソース情報
ヒト・動物細胞 HL-60(RCB3683) RAW 264(RCB0535)