マッシュルーム由来の4-アミノ安息香酸ヒドロキシラーゼに関する研究

概要

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4-Aminobenzoate hydroxylase(4ABH) is a FAD-dependent monooxygenase that catelyzes the decarboxylative hydroxylation of 4-aminobenzoate with the consumption of NAD(P)H and O_2. 4ABH was shown to utilize the proA hydrogen of NAD(P)H. The fact supports the idea that all of the external monoxygenases are involved in the transfer of the proA hydrogen atom of the nicotinamide nucleotide. In order to investigate the structural characteristics of 4ABH, four monoclonal antibodies (mAbs) against the enzyme were produced. Of the mAbs, three mAbs were shown to recognize the FAD-binding domain of the enzyme. The order of the epitopes to the four mAbs was determined by analyzing the immunoblotting patterns of the peptides obtained from the enzyme. The findings provide the evidence for the location of the FAD-binding domain of the enzyme. Interestingly, one of the mAbs recognizing the FAD-binding domain of 4ABH was shown to recognize those of salicylate hydroxylase and _D-amino acid oxidase, supporting the Rossman's hypothesis. The sequence of 37 N-terminal amino acid residues of the peptide fragments obtained by digestion with lysyl endopeptidase has been determined. The cDNA coding the 37 amino acid residues was produced by PCR. Using the probe prepared by random-priming of the cDNA, the mRNA of the enzyme was detected and its size was shown to be approximately 1.7 kb.
日本ビタミン学会の論文
1993-11-25