Novel development and applications of the reagents for chemical scission of targetted proteins as chemical scissors

• Project/Area Number: 25650038
• Research Category: Grant-in-Aid for Challenging Exploratory Research
• Allocation Type: Multi-year Fund
• Research Field: Functional biochemistry
• Research Institution: Kinki University
• Principal Investigator: NOZAKI Osamu 近畿大学, 医学部附属病院, 講師 (50164687)
• Co-Investigator(Kenkyū-buntansha): SHIZUMA Motohiro 地方独立行政法人・大阪市立工業研究所, 生物・生活材料研究部, 研究員 (40416318)
• Research Collaborator: KAWAMOTO Hiroko
• Project Period (FY): 2013-04-01 – 2015-03-31
• Project Status: Completed (Fiscal Year 2014)
• Budget Amount: ¥4,160,000 (Direct Cost: ¥3,200,000、Indirect Cost: ¥960,000); Fiscal Year 2014: ¥2,080,000 (Direct Cost: ¥1,600,000、Indirect Cost: ¥480,000); Fiscal Year 2013: ¥2,080,000 (Direct Cost: ¥1,600,000、Indirect Cost: ¥480,000)
• Keywords: 蛋白切除 / 蛋白過酸化イソインドール / 化学発光 / 蛍光画像測定 / 質量解析 / 試薬開発 / 血清蛋白分画 / o-phthalaldehyde / 蛋白・ペプチド・アミノ酸 / 化学的切除 / Chemical scission / 過酸化イソインドール / 化学発光測定 / 蛍光画像撮影 / 質量分析 / 創薬 / アルブミン / ペプチド / アミノ酸 / chemical scission / αー炭素 / 窒素含有複素環 / オルトフタルアルデヒド蛍光化・過酸化二重修飾

Outline of Final Research Achievements

Development of the novel reagent for chemical scission of targeted proteins and its clinical application　were studied. The reagents those demolish proteins under mild chemical conditions have been requested for treatment of patients and research of diseases. The reagents were protein peroxyisoindoles of proteins, and were prepared by the reaction with hydrogen peroxide following to the reaction with o-phthalaldehyde and thiol compounds. The production of protein peroxyisoindole were confirmed by fluorescence imaging. The peroxyisoindole was oxidized to emit light, accompanying of disappearance of the fluorescence image. This meant demolition of proteins. The reagent of peroxyisoindole was also applied to serum protein fractionated by electrophoresis resulting in successful cleavage. In conclusion, the novel reagents of proteins allowed simultaneous protein scission and monitoring of the accompanying chemiluminescence.

Research Products

• [Journal Article] Chemical scission of albumines, peptides and amino acids and accompanying chemiluminescence after peroxidation with hydrogen peroxide (2014)
  • Author(s): Osamu Nozaki, Motohiro Shizuma
  • Journal Title: Luminescence Volume: 29 Pages: 37-38
  • Peer Reviewed
• [Journal Article] アミノ酸のo-phthalaldehyde・過酸化二重修飾による化学発光測定 (2014)
  • Author(s): 野崎 修、静間基博
  • Journal Title: 日本分析化学会第63年会講演要旨集 Volume: １ Pages: 39-39
• [Presentation] アミノ酸のo-phthalaldehyde・過酸化二重修飾による化学発光測定 (2014)
  • Author(s): 野崎 修、静間基博
  • Organizer: 日本分析化学会第63年会
  • Place of Presentation: 広島大学（東広島市）
  • Year and Date: 2014-09-17 – 2014-09-19
• [Presentation] Chemical scission of albumines, peptides and amino acids and accompanying chemiluminescence after peroxidation with hydrogen peroxide (2014)
  • Author(s): Osamu Nozaki, Motohiro Shizuma
  • Organizer: The 18th International symposium for bioluminescence and chemiluminescence
  • Place of Presentation: Uppsala, Sweden
  • Year and Date: 2014-06-23 – 2014-06-28
• [Presentation] Chemical scission of albumins, peptides and amino acids and accompanying chemiluminescence after peroxidation with hydrogen peroxide (2014)
  • Author(s): Osamu Nozaki, Motohiro Shizuma
  • Organizer: The 18th International Symposium on Bioluminescence and Chemiluminescence
  • Place of Presentation: Uppsala, Sweden.