Development of a method for real-time measuring ion concentrations within membrane contact sites

2015 Fiscal Year Final Research Report

• Project/Area Number: 15K14488
• Research Category: Grant-in-Aid for Challenging Exploratory Research
• Allocation Type: Multi-year Fund
• Research Field: Biophysics
• Research Institution: Osaka University
• Principal Investigator: MAEDA YUSUKE 大阪大学, 微生物病研究所, 准教授 (00294124)
• Project Period (FY): 2015-04-01 – 2016-03-31
• Keywords: 膜接触部位 / pH / カルシウム / バイオイメージング

Outline of Final Research Achievements

To reveal the communications between organelles through membrane contact sites (MCS), we established the assay system to measure ion concentrations within the MCS in real-time as the following. First, we created organelle-specifically localized reporter molecules harboring FRB or FKBP, and these molecules could form MCS by adding rapamycin. Taken pH-resistance into consideration, we created a fusion molecule of GFP and aequorin as a calcium sensor. By fusing organelle-specific reporter molecules with ion-sensitive fluorescent reporter molecules such as pHluorin and calcium sensor, assay systems for ion measurement in MCS were established.

Free Research Field

分子細胞生化学