Proteome analysis and living cell imaging of mast cell secretory granules

Research Project

Project/Area Number	17K08890
Research Category	Grant-in-Aid for Scientific Research (C)
Allocation Type	Multi-year Fund
Section	一般
Research Field	Immunology
Research Institution	Tokyo Women's Medical University
Principal Investigator	Tanaka Shotaro 東京女子医科大学, 医学部, 助教 (90380667)
Project Period (FY)	2017-04-01 – 2020-03-31
Project Status	Completed (Fiscal Year 2019)
Budget Amount *help	¥4,680,000 (Direct Cost: ¥3,600,000、Indirect Cost: ¥1,080,000) Fiscal Year 2019: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000) Fiscal Year 2018: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000) Fiscal Year 2017: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000)
Keywords	オルガネラ調製 / プロテオミクス / マスト細胞 / 分泌顆粒 / 磁性ビーズ / 新規分泌顆粒局在タンパク質 / プロテオーム解析 / セルソーター / アレルギー・ぜんそく / イメージング
Outline of Final Research Achievements	Mast cells are immune cell exist in systemic tissues for innate immune response. Mast cells normally maintain about 1000 of small vesicles named secretory granule (SG) in its cytoplasm which contain various allergy symptom-inducible compounds such as histamine. Details of the mechanism of SG secretion are still controversial because of difficulty to prepare SGs in adequate amount/purity for identification of SG-specific functional molecules. To conquer this problem, we developed a novel method utilizing immune-globulin and magnetic micro-beads to prepare pure SGs with enough amount from mast cells. In consequence, we identified 200 of SG specific proteins from the purified SGs by mass spectrometry. Elucidation of the function of each gene in SG secretion may provide alternate idea to treat allergy symptoms effectively in future.
Academic Significance and Societal Importance of the Research Achievements	アレルギー症状誘発物質の分泌メカニズムの詳細が明らかでなかったのは、純度の高い分泌顆粒を十分量調製するのが難しく、それに係るタンパク質の同定が進んでいないことが原因であった。研究代表者は抗体を連結した磁性ビーズを利用することで、効率の良い分泌顆粒調製技術の開発に成功した。これによって、分泌に係るタンパク質を特定するための道筋ができた。これによって、分泌のメカニズムが解明されるとともに、花粉症やハウスダストによる喘息、食品アレルギーのアレルギー症状を抑制する治療法の開発に貢献できるだろう。