title: Elementary tetrahelical protein design for diverse oxidoreductase functions.
abstract: Emulating functions of natural enzymes in man-made constructs has proven challenging. Here we describe a man-made protein platform that reproduces many of the diverse functions of natural oxidoreductases without importing the complex and obscure interactions common to natural proteins. Our design is founded on an elementary, structurally stable 4-α-helix protein monomer with a minimalist interior malleable enough to accommodate various light- and redox-active cofactors and with an exterior tolerating extensive charge patterning for modulation of redox cofactor potentials and environmental interactions. Despite its modest size, the construct offers several independent domains for functional engineering that targets diverse natural activities, including dioxygen binding and superoxide and peroxide generation, interprotein electron transfer to natural cytochrome c and light-activated intraprotein energy transfer and charge separation approximating the core reactions of photosynthesis, cryptochrome and photolyase. The highly stable, readily expressible and biocompatible characteristics of these open-ended designs promise development of practical in vitro and in vivo applications.
authors.affiliation: Department of Biochemistry and Biophysics, Johnson Research Foundation, University of Pennsylvania, Philadelphia, Pennsylvania, USA.
authors.forename: Tammer A
authors.initials: TA
authors.lastname: Farid
authors.affiliation: Department of Biochemistry and Biophysics, Johnson Research Foundation, University of Pennsylvania, Philadelphia, Pennsylvania, USA.
authors.forename: Goutham
authors.initials: G
authors.lastname: Kodali
authors.affiliation: Department of Biochemistry and Biophysics, Johnson Research Foundation, University of Pennsylvania, Philadelphia, Pennsylvania, USA.
authors.forename: Lee A
authors.initials: LA
authors.lastname: Solomon
authors.affiliation: Department of Biochemistry and Biophysics, Johnson Research Foundation, University of Pennsylvania, Philadelphia, Pennsylvania, USA.|Max Planck Institute for Developmental Biology, Tübingen, Germany.
authors.forename: Bruce R
authors.initials: BR
authors.lastname: Lichtenstein
authors.affiliation: Department of Biochemistry and Biophysics, Johnson Research Foundation, University of Pennsylvania, Philadelphia, Pennsylvania, USA.
authors.forename: Molly M
authors.initials: MM
authors.lastname: Sheehan
authors.affiliation: Department of Biochemistry and Biophysics, Johnson Research Foundation, University of Pennsylvania, Philadelphia, Pennsylvania, USA.
authors.forename: Bryan A
authors.initials: BA
authors.lastname: Fry
authors.affiliation: Department of Biochemistry and Biophysics, Johnson Research Foundation, University of Pennsylvania, Philadelphia, Pennsylvania, USA.
authors.forename: Chris
authors.initials: C
authors.lastname: Bialas
authors.affiliation: Department of Biochemistry and Biophysics, Johnson Research Foundation, University of Pennsylvania, Philadelphia, Pennsylvania, USA.
authors.forename: Nathan M
authors.initials: NM
authors.lastname: Ennist
authors.affiliation: Department of Biochemistry and Biophysics, Johnson Research Foundation, University of Pennsylvania, Philadelphia, Pennsylvania, USA.
authors.forename: Jessica A
authors.initials: JA
authors.lastname: Siedlecki
authors.affiliation: Department of Biochemistry and Biophysics, Johnson Research Foundation, University of Pennsylvania, Philadelphia, Pennsylvania, USA.
authors.forename: Zhenyu
authors.initials: Z
authors.lastname: Zhao
authors.affiliation: Department of Biochemistry and Biophysics, Johnson Research Foundation, University of Pennsylvania, Philadelphia, Pennsylvania, USA.
authors.forename: Matthew A
authors.initials: MA
authors.lastname: Stetz
authors.affiliation: Department of Biochemistry and Biophysics, Johnson Research Foundation, University of Pennsylvania, Philadelphia, Pennsylvania, USA.
authors.forename: Kathleen G
authors.initials: KG
authors.lastname: Valentine
authors.affiliation: Department of Biochemistry and Biophysics, Johnson Research Foundation, University of Pennsylvania, Philadelphia, Pennsylvania, USA.|School of Biochemistry, University of Bristol, Bristol, UK.
authors.forename: J L Ross
authors.initials: JLR
authors.lastname: Anderson
authors.affiliation: Department of Biochemistry and Biophysics, Johnson Research Foundation, University of Pennsylvania, Philadelphia, Pennsylvania, USA.
authors.forename: A Joshua
authors.initials: AJ
authors.lastname: Wand
authors.affiliation: Department of Biochemistry and Biophysics, Johnson Research Foundation, University of Pennsylvania, Philadelphia, Pennsylvania, USA.
authors.forename: Bohdana M
authors.initials: BM
authors.lastname: Discher
authors.affiliation: Department of Biochemistry and Biophysics, Johnson Research Foundation, University of Pennsylvania, Philadelphia, Pennsylvania, USA.
authors.forename: Christopher C
authors.initials: CC
authors.lastname: Moser
authors.affiliation: Department of Biochemistry and Biophysics, Johnson Research Foundation, University of Pennsylvania, Philadelphia, Pennsylvania, USA.
authors.forename: P Leslie
authors.initials: PL
authors.lastname: Dutton
mesh_terms: D006418:Heme / Q000737:chemistry / Q000378:metabolism; D008958:Models, Molecular; D015394:Molecular Structure; D019906:Nuclear Magnetic Resonance, Biomolecular; D010088:Oxidoreductases / Q000737:chemistry / Q000378:metabolism*; D011485:Protein Binding; D011487:Protein Conformation; D015202:Protein Engineering / Q000379:methods; D011506:Proteins / Q000737:chemistry*
chemical_list: D011506:Proteins; D006418:Heme; D010088:Oxidoreductases