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Monitoring Protein Interactions in the Living Cell Through the Fluorescence Decays of the Cyan Fluorescent Protein by Regis Grailhe; Fabienne Merola; Jacqueline Ridard; Stephen Couvignou; Chantal Le Poupon; Jean-Pierre Changeux; Helene Laguitton-Pasquier is a Chemistry article available to read on EtoBox.
What is Monitoring Protein Interactions in the Living Cell Through the Fluorescence Decays of the Cyan Fluorescent Protein about?
## Abstract Using fluorescence lifetime microspectroscopy and imaging techniques, we have studied the fluorescence of cyan fluorescent protein (CFP) transiently expressed in HEK‐293 cells, in the presence or absence of its fluorescence resonance energy transfer (FRET) partner, yellow fluorescent protein (YFP). When the two proteins are attached through a 27‐amino‐acid linker, a 33 % average efficiency of intramolecular energy transfer is accurately determined inside the cell. Additionally, we observe a systematic quenching of the CFP fluorescence with increasing levels of protein expression. This quenching cannot be accounted for by formation of the previously described dimer of GFP‐related proteins, since its magnitude is unchanged when the fluorescent proteins carry the mutation A206K shown to dissociate this dimer in vitro. Even when the intracellular protein concentration largely exceeds the in vitro dissociation constant of the dimer, self‐association remains undetectable, either between free proteins or intramolecularly within the CFP–YFP construct. Instead, the detailed concentration effects are satisfactorily accounted for by a model of intermolecular, concentration‐depende
Who reads Monitoring Protein Interactions in the Living Cell Through the Fluorescence Decays of the Cyan Fluorescent Protein?
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- Author
- Regis Grailhe; Fabienne Merola; Jacqueline Ridard; Stephen Couvignou; Chantal Le Poupon; Jean-Pierre Changeux; Helene Laguitton-Pasquier
- Publisher
- John Wiley and Sons; Wiley (John Wiley & Sons); John Wiley & Sons Ltd.; Wiley (ISSN 1439-4235)
- Published
- 2006
- Language
- EN
- Field
- Chemistry (Physical Sciences)