Evaluation of intracellular GPCR activation by using FRET imaging

2015 Fiscal Year Final Research Report

• Project/Area Number: 26860170
• Research Category: Grant-in-Aid for Young Scientists (B)
• Allocation Type: Multi-year Fund
• Research Field: General pharmacology
• Research Institution: Asahikawa Medical College
• Principal Investigator: Uwada Junsuke 旭川医科大学, 医学部, 助教 (70580314)
• Project Period (FY): 2014-04-01 – 2016-03-31
• Keywords: ムスカリン受容体 / FRET / GPCR

Outline of Final Research Achievements

This research was aimed at elucidating the mechanism of activation of intracellular muscarinic M1 receptor using real-time visualization technique. Twelve FRET constructs of M1 receptor were designed and introduced to N1E-115 neuroblastoma cells. Effectiveness of each constructs is currently being examined using confocal microscopy. In addition, interaction of M1 receptor and associated G proteins were also examined in living cells. Interestingly, Gγ proteins was colocalized with M1 receptors at intracellular vesicles as well as at cell surface. On the other hand, C-terminal tail modified M1 receptor, which is exclusively localized intracellularly, showed poor colocalization with Gγ protein. Therefore, it was indicated that the coupling of intracellular M1 receptor and G proteins is dependent on the trafficking machinery of M1 receptors.

Free Research Field

分子薬理学