TY - JOUR
T1 - Design of an artificial light-harvesting unit by protein engineering
T2 - Cytochrome b562-green fluorescent protein chimera
AU - Takeda, Shuji
AU - Kamiya, Noriho
AU - Arai, Ryoichi
AU - Nagamune, Teruyuki
N1 - Funding Information:
We are grateful to Professor S. G. Sligar for his kind gift of the plasmid-encoding cytb562 gene. Dr. Hiroharu Yui of University of Tokyo is appreciated for his kind assistance in the fluorescence lifetime measurements. We also thank Dr. Mikio Hoshino of RIKEN Institute and Drs. Hiroaki Yonemura and Takashi Hayashi of Kyushu University for helpful discussions. R.A. was supported by JSPS Research Fellowships for Young Scientists.
PY - 2001/11/23
Y1 - 2001/11/23
N2 - We have generated a novel model protein for an artificial light-harvesting complex composed of two proteins, cytochrome b562 (cytb562) and enhanced green fluorescent protein (EGFP), in which two chromophores are fixed in each protein matrix. Cytb562 was appended to the N-terminus of EGFP via a Gly-Ser linker and the resultant fusion protein was successfully expressed in Escherichia coli as a mixture of the apo- and the holo-forms as to the cytb562 moiety. The fluorescence of EGFP was substantially quenched when the apo-form was reconstituted with hemin. Based on the fluorescence lifetime measurements, it appeared that light energy entrapped by EGFP is transferred to the heme of cytb562 by resonance energy transfer (energy transfer yield: 65%). Spatial organization of two chromophores using small and stable protein matrices will be promising toward the construction of an artificial light-harvesting complex by protein engineering.
AB - We have generated a novel model protein for an artificial light-harvesting complex composed of two proteins, cytochrome b562 (cytb562) and enhanced green fluorescent protein (EGFP), in which two chromophores are fixed in each protein matrix. Cytb562 was appended to the N-terminus of EGFP via a Gly-Ser linker and the resultant fusion protein was successfully expressed in Escherichia coli as a mixture of the apo- and the holo-forms as to the cytb562 moiety. The fluorescence of EGFP was substantially quenched when the apo-form was reconstituted with hemin. Based on the fluorescence lifetime measurements, it appeared that light energy entrapped by EGFP is transferred to the heme of cytb562 by resonance energy transfer (energy transfer yield: 65%). Spatial organization of two chromophores using small and stable protein matrices will be promising toward the construction of an artificial light-harvesting complex by protein engineering.
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U2 - 10.1006/bbrc.2001.5966
DO - 10.1006/bbrc.2001.5966
M3 - Article
C2 - 11708816
AN - SCOPUS:0035940926
VL - 289
SP - 299
EP - 304
JO - Biochemical and Biophysical Research Communications
JF - Biochemical and Biophysical Research Communications
SN - 0006-291X
IS - 1
ER -