Structural characterization of the photoswitchable fluorescent protein Dronpa-C62S.
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 Angstroms and 1.8 Angstroms, 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 homologs.
Nam
Ki-Hyun
KH
Division of Biotechnology, College of Life Sciences & Biotechnology, Korea University, Anam-Dong, Sungbuk-Gu, Seoul, South Korea.
Kwon
Oh Yeun
OY
Sugiyama
Kanako
K
Lee
Won-Ho
WH
Kim
Young Kwan
YK
Song
Hyun Kyu
HK
Kim
Eunice Eunkyung
EE
Park
Sam-Yong
SY
Jeon
Hyesung
H
Hwang
Kwang Yeon
KY
D000818:Animals; D037421:Anthozoa; D003460:Crystallization; D018360:Crystallography, X-Ray; D049452:Green Fluorescent Proteins / Q000737:chemistry* / Q000235:genetics; D006860:Hydrogen Bonding; D008958:Models, Molecular
D049452:Green Fluorescent Proteins