Multi-step excitation energy transfer engineered in genetic fusions of natural and synthetic light-harvesting proteins.
Synthetic proteins designed and constructed from first principles with minimal reference to the sequence of any natural protein have proven robust and extraordinarily adaptable for engineering a range of functions. Here for the first time we describe the expression and genetic fusion of a natural photosynthetic light-harvesting subunit with a synthetic protein designed for light energy capture and multi-step transfer. We demonstrate excitation energy transfer from the bilin of the CpcA subunit (phycocyanin α subunit) of the cyanobacterial photosynthetic light-harvesting phycobilisome to synthetic four-helix-bundle proteins accommodating sites that specifically bind a variety of selected photoactive tetrapyrroles positioned to enhance energy transfer by relay. The examination of combinations of different bilin, chlorin and bacteriochlorin cofactors has led to identification of the preconditions for directing energy from the bilin light-harvesting antenna into synthetic protein-cofactor constructs that can be customized for light-activated chemistry in the cell.
Department of Biochemistry and Biophysics, University of Pennsylvania, Philadelphia, PA 19104, USA.
Joshua A
0000-0001-5151-5643
JA
Mancini
Department of Biochemistry and Biophysics, University of Pennsylvania, Philadelphia, PA 19104, USA.
Goutham
0000-0001-6356-9016
G
Kodali
Department of Chemistry, North Carolina State University, Raleigh, NC 27695, USA.
Jianbing
0000-0003-4792-8140
J
Jiang
Department of Chemistry, North Carolina State University, Raleigh, NC 27695, USA.
Kanumuri Ramesh
0000-0002-4795-6937
KR
Reddy
Department of Chemistry, North Carolina State University, Raleigh, NC 27695, USA.
Jonathan S
0000-0002-4872-2040
JS
Lindsey
Department of Biochemistry and Molecular Biology, The Pennsylvania State University, University Park, PA 16802, USA.
Donald A
0000-0003-2782-5988
DA
Bryant
Department of Biochemistry and Biophysics, University of Pennsylvania, Philadelphia, PA 19104, USA.
P Leslie
0000-0002-3063-3154
PL
Dutton
Department of Biochemistry and Biophysics, University of Pennsylvania, Philadelphia, PA 19104, USA moserc@mail.med.upenn.edu.
Christopher C
0000-0003-4814-8568
CC
Moser
D001426:Bacterial Proteins / Q000737:chemistry* / Q000235:genetics; D010798:Phycocyanin / Q000737:chemistry* / Q000235:genetics; D011166:Porphyrins / Q000737:chemistry* / Q000235:genetics; D046939:Synechocystis / Q000737:chemistry* / Q000235:genetics
D001426:Bacterial Proteins; D011166:Porphyrins; C452433:bacteriochlorin; D010798:Phycocyanin
excitation energy transfer; fusion protein; light-harvesting antenna; maquette; phycobiliprotein; synthetic protein