Protein-ligand complex formation revealed by fluorescence lifetime-correlation analysis

2016 Fiscal Year Final Research Report

• Project/Area Number: 15K17814
• Research Category: Grant-in-Aid for Young Scientists (B)
• Allocation Type: Multi-year Fund
• Research Field: Physical chemistry
• Research Institution: Saitama University
• Principal Investigator: OTOSU Takuhiro 埼玉大学, 研究機構研究企画推進室, 助教 (90564948)
• Project Period (FY): 2015-04-01 – 2017-03-31
• Keywords: 蛍光寿命相関分光 / 蛋白質 / 構造ダイナミクス

Outline of Final Research Achievements

Conformational fluctuations of proteins are highly relevant to their biological functions. To this aim, we developed two-dimensional fluorescence lifetime correlation spectroscopy (2D FLCS) which analyze the conformational dynamics of biomolecules in an equilibrium by the correlation analysis of fluorescence lifetime. The aim of this study is to extend 2D FLCS to analyze the conformational dynamics in a non-equilibrium condition. For this purpose, I developed new 2D FLCS instrument which utilizes two laser pulses (UV pulse and visible pulse). UV pulse is used for uncaging the caged compound to transiently increase the ligand concentration in the focal region. The conformational change of protein caused by the interaction with the ligand is then monitored through visible pulse. Preliminary results clearly showed that the instrument developed here is applicable to analyze the ligand-induced conformational changes of proteins with 2D FLCS.

Free Research Field

生物物理