Document Detail

Reversible single-molecule photoswitching in the GFP-like fluorescent protein Dronpa.
MedLine Citation:
PMID:  15972810     Owner:  NLM     Status:  MEDLINE    
Reversible photoswitching of individual molecules has been demonstrated for a number of mutants of the green fluorescent protein (GFP). To date, however, a limited number of switching events with slow response to light have been achieved at the single-molecule level. Here, we report reversible photoswitching characteristics observed in individual molecules of Dronpa, a mutant of a GFP-like fluorescent protein that was cloned from a coral Pectiniidae. Ensemble spectroscopy shows that intense irradiation at 488 nm changes Dronpa to a dim protonated form, but even weak irradiation at 405 nm restores it to the bright deprotonated form. Although Dronpa exists in an acid-base equilibrium, only the photoinduced protonated form shows the switching behavior. At the single-molecule level, 488- and 405-nm lights can be used to drive the molecule back and forth between the bright and dim states. Such reversible photoswitching could be repeated >100 times. The response speed to irradiation depends almost linearly on the irradiation power, with the response time being in the order of milliseconds. The perfect reversibility of the Dronpa photoswitching allows us to propose a detailed model, which quantitatively describes interconversion among the various states. The fast response of Dronpa to light holds great promise for following fast diffusion or transport of signaling molecules in live cells.
Satoshi Habuchi; Ryoko Ando; Peter Dedecker; Wendy Verheijen; Hideaki Mizuno; Atsushi Miyawaki; Johan Hofkens
Publication Detail:
Type:  Comparative Study; Journal Article; Research Support, Non-U.S. Gov't     Date:  2005-06-22
Journal Detail:
Title:  Proceedings of the National Academy of Sciences of the United States of America     Volume:  102     ISSN:  0027-8424     ISO Abbreviation:  Proc. Natl. Acad. Sci. U.S.A.     Publication Date:  2005 Jul 
Date Detail:
Created Date:  2005-07-06     Completed Date:  2005-09-12     Revised Date:  2010-09-21    
Medline Journal Info:
Nlm Unique ID:  7505876     Medline TA:  Proc Natl Acad Sci U S A     Country:  United States    
Other Details:
Languages:  eng     Pagination:  9511-6     Citation Subset:  IM    
Department of Chemistry, Katholieke Universiteit Leuven, Celestijnenlaan 200F, 3001 Heverlee, Belgium.
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MeSH Terms
Anthozoa / chemistry*
Green Fluorescent Proteins / chemistry*,  radiation effects
Models, Chemical*
Spectrometry, Fluorescence / methods
Time Factors
Reg. No./Substance:
147336-22-9/Green Fluorescent Proteins
Comment In:
Proc Natl Acad Sci U S A. 2005 Jul 5;102(27):9433-4   [PMID:  15983383 ]

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