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Title 

Fluorescence lifetime study of ECFP/EYFP labeled MBP in Frster resonance energy transfer

Authors 

S LeeS Y KimKyoung Sook ParkBong Hyun ChungS W Kim

Publisher 

Elsevier

Issue Date 

2011

Citation 

Journal of Luminescence, vol. 131, no. 2, pp. 275-279

Keywords 

FRETLifetimeMaltose binding proteinTCSPC

Abstract 

This paper studies the conformational change of the binding protein by a fluorescence lifetime method. As a model protein, maltose binding protein (MBP) where enhanced cyan protein (ECFP) and enhanced yellow fluorescent protein (EYFP) were genetically fused to act as a donor and an acceptor in Frster resonance energy transfer (FRET) was used. The ECFP and the EYFP were linked to the C-terminal and N-terminal regions of the MBP, respectively. In order to investigate the conformational change of the MBP, the lifetime of the ECFP, which acts as a donor in the ECFP:MBP:EYFP fusion protein, was analyzed during the FRET process. We observed that two lifetime components exist when the ECFP is linked to the MBP and that the lifetime of the ECFP is shortened when ECFP:MBP:EYFP protein undergoes a conformational change as a result of the maltose binding. In addition, we observed that the lifetime of the donor is gradually shorter in the ECFP:MBP:EYFP fusion protein as the maltose concentration increases. By a lifetime analysis and simulation study, we found that the participant rate of the ECFP:MBP:EYFP protein in FRET is the main cause of the donor lifetime shortening in relation to the increase of the maltose concentration.

ISSN 

0022-2313

Link 

http://dx.doi.org/10.1016/j.jlumin.2010.10.012

Appears in Collections

1. Journal Articles > Journal Articles

Registered Date

2017-04-19


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