De novo synthetic biliprotein design, assembly and excitation energy transfer. Issue 141 (30th April 2018)

• Record Type: Journal Article
• Authors: Mancini, Joshua A.; Sheehan, Molly; Kodali, Goutham; Chow, Brian Y.; Bryant, Donald A.; Dutton, P. Leslie; Moser, Christopher C.
• Is Part Of: Journal of the Royal Society interface. Volume 15:Issue 141(2018)
• Publication Date: 2018-04-30
• Subjects: synthetic protein -- maquette -- biliprotein -- light-harvesting -- excitation energy transfer; Physical sciences -- Research -- Periodicals; Life sciences -- Research -- Periodicals; Interdisciplinary research -- Periodicals; 570.5
• DOI: 10.1098/rsif.2018.0021
• Languages: English
• ISSNs: 1742-5689

Abstract

Abstract : Bilins are linear tetrapyrrole chromophores with a wide range of visible and near-visible light absorption and emission properties. These properties are tuned upon binding to natural proteins and exploited in photosynthetic light-harvesting and non-photosynthetic light-sensitive signalling. These pigmented proteins are now being manipulated to develop fluorescent experimental tools. To engineer the optical properties of bound bilins for specific applications more flexibly, we have used first principles of protein folding to design novel, stable and highly adaptable bilin-binding four-α-helix bundle protein frames, called maquettes, and explored the minimal requirements underlying covalent bilin ligation and conformational restriction responsible for the strong and variable absorption, fluorescence and excitation energy transfer of these proteins. Biliverdin, phycocyanobilin and phycoerythrobilin bind covalently to maquette Cys in vitro . A blue-shifted tripyrrole formed from maquette-bound phycocyanobilin displays a quantum yield of 26%. Although unrelated in fold and sequence to natural phycobiliproteins, bilin lyases nevertheless interact with maquettes during co-expression in Escherichia coli to improve the efficiency of bilin binding and influence bilin structure. Bilins bind in vitro and in vivo to Cys residues placed in loops, towards the amino end or in the middle of helices but bind poorly at the carboxyl end of helices. Bilin-binding efficiency and fluorescence yield are improved by Arg and Asp residues adjacent to the ligating Cys on the same helix and by His residues on adjacent helices.