Proteomic analysis of IgA-deposits on glomeruli of biopsy samples of IgA nephropathy

2015 Fiscal Year Final Research Report

• Project/Area Number: 25461212
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Kidney internal medicine
• Research Institution: Niigata University
• Principal Investigator: Yoshida Yutaka 新潟大学, 研究推進機構, 特任専門職員 (40182795)
• Co-Investigator(Kenkyū-buntansha): YAOITA Eishin 新潟大学, 大学院医歯学総合研究科, 准教授 (00157950) ; YAMAMOTO Tadashi 新潟大学, 産学地域連携推進機構, 特任教授 (30092737)
• Project Period (FY): 2013-04-01 – 2016-03-31
• Keywords: IgA腎症 / IgA免疫複合体 / 凍結腎生検標本 / 単離糸球体 / スリット膜 / EMARS法 / プロテオミクス

Outline of Final Research Achievements

EMARS method can label proteins in the immediate vicinity of a target membrane protein with fluorescein without destruction of cells or tissue integrity. We aimed to identify IgA deposits on glomerulus of biopsy samples of patients with IgA nephropathy bur found problems in specificity of labeling and low recovery of proteins. We, therefore, used glomeruli highly purified from rat kidneys and nephrin as a target protein to improve EMARS method. By using fluorescein-tyramide as a labeling reagent, labeling specificity was much improved and we succeeded in identification of many membrane proteins associates with nephrin in the slit diaphragm, but also find a low recovery and low specificity of labeled proteins in the step of collection of labeled proteins.

Free Research Field

腎糸球体のプロテオミクス