title: Increased Diels-Alderase activity through backbone remodeling guided by Foldit players.
abstract: Computational enzyme design holds promise for the production of renewable fuels, drugs and chemicals. De novo enzyme design has generated catalysts for several reactions, but with lower catalytic efficiencies than naturally occurring enzymes. Here we report the use of game-driven crowdsourcing to enhance the activity of a computationally designed enzyme through the functional remodeling of its structure. Players of the online game Foldit were challenged to remodel the backbone of a computationally designed bimolecular Diels-Alderase to enable additional interactions with substrates. Several iterations of design and characterization generated a 24-residue helix-turn-helix motif, including a 13-residue insertion, that increased enzyme activity >18-fold. X-ray crystallography showed that the large insertion adopts a helix-turn-helix structure positioned as in the Foldit model. These results demonstrate that human creativity can extend beyond the macroscopic challenges encountered in everyday life to molecular-scale design problems.
authors.affiliation: Department of Biochemistry, University of Washington, Seattle, Washington, USA.
authors.forename: Christopher B
authors.initials: CB
authors.lastname: Eiben
authors.forename: Justin B
authors.initials: JB
authors.lastname: Siegel
authors.forename: Jacob B
authors.initials: JB
authors.lastname: Bale
authors.forename: Seth
authors.initials: S
authors.lastname: Cooper
authors.forename: Firas
authors.initials: F
authors.lastname: Khatib
authors.forename: Betty W
authors.initials: BW
authors.lastname: Shen
authors.forename: Foldit
authors.initials: F
authors.lastname: Players
authors.forename: Barry L
authors.initials: BL
authors.lastname: Stoddard
authors.forename: Zoran
authors.initials: Z
authors.lastname: Popovic
authors.forename: David
authors.initials: D
authors.lastname: Baker
mesh_terms: D000465:Algorithms; D002384:Catalysis; D019295:Computational Biology; D018360:Crystallography, X-Ray; D004798:Enzymes / Q000138:chemical synthesis* / Q000737:chemistry*; D019077:Helix-Turn-Helix Motifs; D006801:Humans; D008958:Models, Molecular; D015202:Protein Engineering / Q000379:methods*; D013329:Structure-Activity Relationship; D013379:Substrate Specificity; D018910:Video Games
chemical_list: D004798:Enzymes