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Biophys. J. BioFAST: First Published June 23, 2006. doi:10.1529/biophysj.106.089789
© 2006 by the Biophysical Society.

A more recent version of this article appeared on September 1, 2006.
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BIOPHYSICAL LETTERS

Fast and Reversible Photoswitching of the Fluorescent Protein Dronpa as Evidenced by Fluorescence Correlation Spectroscopy

Peter Dedecker 1, Jun-ichi Hotta 1, Ryoko Ando 2, Atsushi Miyawaki 2, Yves Engelborghs 3 and Johan Hofkens 1*

1 Katholieke Universiteit Leuven
2 RIKEN
3 K.U. Leuven

* To whom correspondence should be addressed. E-mail: johan.hofkens{at}chem.kuleuven.be.

Submitted on May 22, 2006
Revised on June 8, 2006
Accepted on 16 June 2006


  Abstract
Controlling molecular properties through photoirradiation holds great promise for its potential for non-invasive and selective manipulation of matter. Photochromism has been observed for several different molecules, including green fluorescent proteins, and recently the discovery of a novel photoswitchable green fluorescent protein called Dronpa was reported. Dronpa displays reversible and highly efficient on/off photoswitching of its fluorescence emission, and reversible switching of immobilized single molecules of Dronpa with response times faster than 20 milliseconds was demonstrated. In this letter we expand these observations to freely diffusing molecules by using fluorescence correlation spectroscopy with simultaneous excitation at 488 and 405 nm. By varying the intensity of irradiation at 405 nm, we demonstrate the reversible photoswitching of Dronpa under these conditions, and from the obtained autocorrelation functions we conclude that this photoswitching can occur within tens of microseconds.

Key Words: Dronpa, Fluorescence correlation spectroscopy, Green fluorescent protein, Photoswitching, Single-molecule




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Copyright © 2006 by the Biophysical Society.