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De novo synthetic biliprotein design, assembly and excitation energy transfer.
Mancini, Joshua A; Sheehan, Molly; Kodali, Goutham; Chow, Brian Y; Bryant, Donald A; Dutton, P Leslie; Moser, Christopher C.
Afiliación
  • Mancini JA; Department of Biochemistry and Biophysics, University of Pennsylvania, Philadelphia, PA, USA.
  • Sheehan M; Department of Bioengineering, University of Pennsylvania, Philadelphia, PA, USA.
  • Kodali G; Department of Biochemistry and Biophysics, University of Pennsylvania, Philadelphia, PA, USA.
  • Chow BY; Department of Bioengineering, University of Pennsylvania, Philadelphia, PA, USA.
  • Bryant DA; Department of Biochemistry and Molecular Biology, The Pennsylvania State University, University Park, PA, USA.
  • Dutton PL; Department of Biochemistry and Biophysics, University of Pennsylvania, Philadelphia, PA, USA.
  • Moser CC; Department of Biochemistry and Biophysics, University of Pennsylvania, Philadelphia, PA, USA moserc@mail.med.upenn.edu.
J R Soc Interface ; 15(141)2018 04.
Article en En | MEDLINE | ID: mdl-29618529
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.
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Transferencia de Energía / Ficobiliproteínas Tipo de estudio: Prognostic_studies Idioma: En Revista: J R Soc Interface Año: 2018 Tipo del documento: Article País de afiliación: Estados Unidos Pais de publicación: Reino Unido

Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Transferencia de Energía / Ficobiliproteínas Tipo de estudio: Prognostic_studies Idioma: En Revista: J R Soc Interface Año: 2018 Tipo del documento: Article País de afiliación: Estados Unidos Pais de publicación: Reino Unido