Rational design of a protein-based molecular device consisting of blue fluorescent protein and zinc protoporphyrin IX incorporated into a cytochrome b562 scaffold.
To make a single molecular photo-device, it is essential to control the exact orientation of two types of proteins. We made a chimeric protein in which cytochrome b562 was linked to the N-terminus of enhanced green fluorescent protein, cytb562-EGFP. Within cytb562-EGFP, the excitation energy of EGFP was transferred to the cytochrome b562 cofactor fixed proximally to EGFP. Cytb562-EGFP was engineered so that iron protoporphyrin IX was substituted by zinc protoporphyrin IX to make it a suitable cofactor for photo-induced electron transfer. The photosensitizer pigment was optimized and the EGFP was replaced by a blue fluorescent mutant that gave 15% higher energy transfer efficiency.
Department of Chemistry and Biotechnology, School of Engineering, The University of Tokyo, 7-3-1 Hongo Bunkyo-ku, Tokyo 113-8656, Japan.
Shuji
S
Takeda
Noriho
N
Kamiya
Teruyuki
T
Nagamune
D003573:Cytochrome b Group / Q000737:chemistry*; D004735:Energy Transfer; D029968:Escherichia coli Proteins / Q000737:chemistry*; D049452:Green Fluorescent Proteins; D008164:Luminescent Proteins / Q000737:chemistry*; D008958:Models, Molecular; D011487:Protein Conformation; D015202:Protein Engineering; D011524:Protoporphyrins / Q000737:chemistry*; D011993:Recombinant Fusion Proteins / Q000737:chemistry; D013050:Spectrometry, Fluorescence
D003573:Cytochrome b Group; D029968:Escherichia coli Proteins; D008164:Luminescent Proteins; D011524:Protoporphyrins; D011993:Recombinant Fusion Proteins; C409504:blue fluorescent protein, Aequorea victoria; C448299:iron protoporphyrin IX; D049452:Green Fluorescent Proteins; C017803:zinc protoporphyrin; C017850:cytochrome b562, E coli