title: Epitope mapping of monoclonal antibodies against 4-aminobenzoate hydroxylase from Agaricus bisporus.
abstract: Four monoclonal antibodies (mAbs) against 4-aminobenzoate hydroxylase (EC 1.14.13.27) have been produced (H. Tsuji et al., J. Biol. Chem. 265 (1990) 16064; T. Ogawa et al., Biochim. Biophys. Acta 1115 (1992) 220). Of the mAbs, three mAbs (mAb-A, -B1 and -B2) recognize the FAD-binding domain of the enzyme. In the present study, the epitopes of the mAbs on the enzyme have been examined using pGEX-2T expression systems for DNA fragments encoding various partial amino acid sequences of 4-aminobenzoate hydroxylase. The epitopes for mAb-A, -B1 and -B2 were shown to be on sequences 413-434, 435-460 and 380-413, respectively. These findings suggest that these epitopes for mAb-A, -B1 and -B2 may be close to the isoalloxazine moiety of FAD, which plays a central role in the catalysis of the enzyme.
authors.affiliation: Department of Nutritional Science, Faculty of health and Welfare Science, Okayama Prefectural University, Soja, Okayama 719-1197, Japan.
authors.forename: H
authors.initials: H
authors.lastname: Tsuji
authors.forename: M
authors.initials: M
authors.lastname: Kimoto
authors.forename: H
authors.initials: H
authors.lastname: Watanabe
authors.forename: T
authors.initials: T
authors.lastname: Sasagawa
authors.forename: T
authors.initials: T
authors.lastname: Oka
authors.forename: H
authors.initials: H
authors.lastname: Yamashita
authors.forename: M
authors.initials: M
authors.lastname: Okita
mesh_terms: D000364:Agaricus / Q000201:enzymology*; D000911:Antibodies, Monoclonal / Q000737:chemistry / Q000276:immunology*; D004247:DNA / Q000096:biosynthesis; D000939:Epitopes / Q000032:analysis*; D004926:Escherichia coli / Q000378:metabolism; D005182:Flavin-Adenine Dinucleotide / Q000276:immunology; D005982:Glutathione Transferase / Q000737:chemistry / Q000276:immunology; D006899:Mixed Function Oxygenases / Q000737:chemistry / Q000276:immunology*; D010957:Plasmids; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000276:immunology*
chemical_list: D000911:Antibodies, Monoclonal; D000939:Epitopes; D011993:Recombinant Fusion Proteins; D005182:Flavin-Adenine Dinucleotide; D004247:DNA; D006899:Mixed Function Oxygenases; C050092:4-aminobenzoate hydroxylase; D005982:Glutathione Transferase