Protection mechanism against bacteria by platelet: A novel strategy for sepsis

2018 Fiscal Year Final Research Report

• Project/Area Number: 16K10957
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Anesthesiology
• Research Institution: Mie University
• Principal Investigator: Kamei Masataka 三重大学, 医学部附属病院, 教授 (60443503)
• Co-Investigator(Kenkyū-buntansha): 島岡 要 三重大学, 医学系研究科, 教授 (40281133)
• Project Period (FY): 2016-04-01 – 2019-03-31
• Keywords: 敗血症 / 血小板 / プロテオミクス

Outline of Final Research Achievements

Optimization of quantitative proteome of blood was performed using ITRAQ method as a pilot study. The iTRAQ method is based on the covalent labeling of the N-terminus and side chain amines of peptides from protein digestions with tags of varying mass. Removal of albumin was needed. The blood proteins without albumin were successfully labeled by iTRAQ in our laboratory, and the samples were then pooled and usually fractionated by liquid chromatography and analyzed by tandem mass spectrometry (MS/MS). A database search is then performed using the fragmentation data to identify the labeled peptides and hence the corresponding proteins.

Free Research Field

麻酔蘇生学

Academic Significance and Societal Importance of the Research Achievements

血液検体から抽出されたタンパクをアルブミン除去後にiTRAQ標識することに成功したことで、今後の血液検体の定量的プロテオミクスの道筋ができた。アルブミン除去処置をした後のタンパクサンプルのiTRAQ標識にて、100個ほどのペプチド同定、データベースとの照合によるタンパク同定を行うことができた。本研究では血小板タンパクを標的とするが、今回構築された血液検体タンパクのデータベースは、血小板の抽出効率の評価に使用できる。