Development of functional human serum albumin (HSA) mutants and studies of their ABT recognition mechanisms

• Project/Area Number: 24390028
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Partial Multi-year Fund
• Section: 一般
• Research Field: Drug development chemistry
• Research Institution: Kumamoto University
• Principal Investigator: MORIOKA HIROSHI 熊本大学, 大学院生命科学研究部(薬学系), 教授 (20230097)
• Co-Investigator(Kenkyū-buntansha): OTAGIRI Masaki 崇城大学, 薬学部, 教授 (80120145) ; KOBASHIGAWA Yoshihiro 熊本大学, 大学院生命科学研究部, 准教授 (90455600) ; SUWA Yoshiaki 熊本大学, 薬学部, 特任助教 (50516127)
• Project Period (FY): 2012-04-01 – 2015-03-31
• Project Status: Completed (Fiscal Year 2014)
• Budget Amount: ¥18,200,000 (Direct Cost: ¥14,000,000、Indirect Cost: ¥4,200,000); Fiscal Year 2014: ¥3,250,000 (Direct Cost: ¥2,500,000、Indirect Cost: ¥750,000); Fiscal Year 2013: ¥5,460,000 (Direct Cost: ¥4,200,000、Indirect Cost: ¥1,260,000); Fiscal Year 2012: ¥9,490,000 (Direct Cost: ¥7,300,000、Indirect Cost: ¥2,190,000)
• Keywords: 薬物認識 / ヒト血清アルブミン / ビリルビン / タンパク質工学 / 分子進化工学 / ファージディスプレイ / ランダム変異 / ビリルビン排泄促進剤

Outline of Final Research Achievements

We have obtained several recombinant human serum albumin (HSA) domain II mutant proteins with high bilirubin (BR) binding affinity by phage display technology. The BR binding affinity of RWLR mutant was found to be 7 times higher than that of WT (FWRR). The results of docking simulation studies and induced CD spectra analyses of BR bound to HSA domain II protein (WT and RWLR mutant) showed that the entrance of BR-binding pocket in RWLR mutant has expanded by the replacement of Arg with Leu at position 218, suggesting that BR could enter the binding pocket more deeply as compared with WT (FWRR). Administration of RWLR mutant reduced serum BR level and increased its urinary excretion in the disease model mice as compared to WT (FWRR) treatment.

Research Products

• [Journal Article] Albumin domain II mutant with high bilirubin binding affinity has a great potential as serum bilirubin excretion enhancer for hyperbilirubinemia treatment. (2013)
  • Author(s): Minomo A
  • Journal Title: Biochim Biophys Acta. Volume: 印刷中 Issue: 4 Pages: 141-8
  • DOI: 10.1016/j.bbagen.2013.01.006
  • Peer Reviewed
• [Presentation] Albumin domain II mutant with high bilirubin binding affinity has a great potential as serum bilirubin excretion enhancer for hyperbilirubinemia treatment. (2013)
  • Author(s): Toru Maruyama, Yu Ishima, Ai Minomo, Toru Narisoko, Hiroshi Morioka, Hiroshi Watanabe, Masaki Otagiri
  • Organizer: The 5th Asian Arden Conference
  • Place of Presentation: Aichi Gakuin University
• [Presentation] SPRによる自己組織化膜(SAMs)上に固定化されたビリルビンとヒト血清アルブミンドメインの結合評価 (2012)
  • Author(s): 笠毛映理子,蓑毛 藍,堀内智裕,諏訪喜昭,渡辺栄治,異島 優,丸山 徹,小田切優樹,森岡弘志
  • Organizer: BMAS2012 第25回バイオメディカル分析科学シンポジウム
  • Place of Presentation: 慶応義塾大学薬学部 芝共立キャンパス
  • Year and Date: 2012-08-09
• [Book] Advanced Methods in Structural Biology (Surface Plasmon Resonance) (2015)
  • Author(s): Kobashigawa, Y., Fukuda, N., Nakahara, Y., Morioka, H. (Senda, T. and Maenaka, K., eds)
  • Total Pages: 15
  • Publisher: Springer
• [Remarks] 新たな治療法を求めて ～血液透析療法に一石を～
  • URL: http://www.pharm.kumamoto-u.ac.jp/Labs/bunseki/research.html#n33
• [Remarks] ヒト血清アルブミン（HSA）の生体内動態制御機構および組換え型HSA製剤の臨床応用に関する研究
  • URL: http://www.pharm.kumamoto-u.ac.jp/Labs/Yakuzai/research/index.html
• [Remarks] 新たな抗体分子をつくり医療につなげよう ～新薬を創る・病気の状態を調べる・治療法を開発する～
  • URL: http://www.pharm.kumamoto-u.ac.jp/center/souyaku/project/morioka.html
• [Remarks] 新たな抗体分子をつくり医療につなげよう ～新薬を創る・病気の状態を調べる・治療法を開発する～
  • URL: http://www.pharm.kumamoto-u.ac.jp/center/souyaku/project/morioka.html