Structural characterization of the photoswitchable fluorescent protein Dronpa-C62S

Ki Hyun Nam, Oh Yeun Kwon, Kanako Sugiyama, Won Ho Lee, Young Kwan Kim, Hyun Kyu Song, Eunice Eunkyung Kim, Sam Yong Park, Hyesung Jeon, Kwang Yeon Hwang

Research output: Contribution to journalArticle

19 Citations (Scopus)

Abstract

The photoswitching behavior of green fluorescent proteins (GFPs) or GFP-like proteins is increasingly recognized as a new technique for optical marking. Recently, Ando and his colleagues developed a new green fluorescent protein Dronpa, which possesses the unique photochromic property of being photoswitchable in a non-destructive manner. To better understand this mechanism, we determined the crystal structures of a new GFP Dronpa and its mutant C62S, at 1.9 Å and 1.8 Å, respectively. Determination of the structures demonstrates that a unique hydrogen-bonding network and the sulfur atom of the chromophore are critical to the photoswitching property of Dronpa. Reversible photoswitching was lost in cells expressing the Dronpa-C62S upon repetitive irradiation compared to the native protein. Structural and mutational analyses reveal the chemical basis for the functional properties of photoswitchable fluorescent proteins and provide the basis for subsequent coherent engineering of this subfamily of Dronpa homolog's.

Original languageEnglish
Pages (from-to)962-967
Number of pages6
JournalBiochemical and biophysical research communications
Volume354
Issue number4
DOIs
Publication statusPublished - 2007 Mar 23

    Fingerprint

Keywords

  • Dronpa
  • Dronpa-C62S
  • Isomerization
  • Photoswitchable
  • Real-time imaging

ASJC Scopus subject areas

  • Biophysics
  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this