P219L substitution in human D-amino acid oxidase impacts the ligand binding and catalytic efficiency

• Material Type: 博士論文
• Title: P219L substitution in human D-amino acid oxidase impacts the ligand binding and catalytic efficiency
• Author/Editor: Rachadech, Wanitcha; Kato, Yusuke; El-Magd, Rabab M. Abou; Shishido, Yuji; Kim, Soo Hyeon; Sogabe, Hirofumi; Maita, Nobuo; Yorita, Kazuko; Fukui, Kiyoshi
• Author Heading: Rachadech, Wanitcha; Kato, Yusuke; El-Magd, Rabab M. Abou; Shishido, Yuji; Kim, Soo Hyeon; Sogabe, Hirofumi; Maita, Nobuo; Yorita, Kazuko; Fukui, Kiyoshi
• Publication, Distribution, etc.: Oxford University Press
• Publication Date: 2020-07-30
• Publication Date (W3CDTF): 2020-07-30
• Alternative Title: ヒトD-アミノ酸酸化酵素のP219L置換が与えるリガンド結合と触媒効率に対する影響; P219L DAO alters ligand binding and catalytic efficiency
• Periodical title: The Journal of Biochemistry
• No. or year of volume/issue: 168 5
• Volume: 168
• Issue: 5
• Pages: 557-567
• ISSN (Periodical Title): ISSN : 1756-2651; ISSN : 0021-924X
• Degree grantor/type: 徳島大学
• Date Granted: 2021-03-23
• Dissertation Number: 甲第3497号
• Degree Type: 博士（医学）
• Text Language Code: eng
• Subject Heading: human D-amino acid oxidase; point-mutation; active site lid; structure–function relationship; X-ray crystallography
• Target Audience: 一般
• Note (General): Human D-amino acid oxidase (DAO) is a flavoenzyme that is implicated in neurodegenerative diseases. We investigated the impact of replacement of proline with leucine at position 219 (P219L) in the active site lid of human DAO on the structural and enzymatic properties, because porcine DAO contains leucine at the corresponding position. The turnover numbers (kcat) of P219L were unchanged, but its Km values decreased compared to wild-type, leading to an increase in the catalytic efficiency (kcat/Km). Moreover, benzoate inhibits P219L with lower Ki value (0.7-0.9 μM) compared to wild-type (1.2-2.0 μM). Crystal structure of P219L in complex with flavin adenine dinucleotide (FAD) and benzoate at 2.25 Å resolution displayed conformational changes of the active site and lid. The distances between the H-bond-forming atoms of arginine 283 and benzoate and the relative position between the aromatic rings of tyrosine 224 and benzoate were changed in the P219L complex. Taken together, the P219L substitution leads to an increase in the catalytic efficiency and binding affinity for substrates/inhibitors due to these structural changes. Furthermore, an acetic acid was located near the adenine ring of FAD in the P219L complex. The present study provides new insights into the structure-function relationship of human DAO.
• Format (IMT): application/pdf
• Access Restrictions: インターネット公開
• Related Material (DOI): 10.1093/jb/mvaa083
• Data Provider (Database): 国立情報学研究所 : 学術機関リポジトリデータベース（IRDB）（機関リポジトリ）; https://irdb.nii.ac.jp
• Original Data Provider (Database): 徳島大学 : 徳島大学機関リポジトリ

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