title: Dynamics and hydration explain failed functional transformation in dehalogenase design.
abstract: We emphasize the importance of dynamics and hydration for enzymatic catalysis and protein design by transplanting the active site from a haloalkane dehalogenase with high enantioselectivity to nonselective dehalogenase. Protein crystallography confirms that the active site geometry of the redesigned dehalogenase matches that of the target, but its enantioselectivity remains low. Time-dependent fluorescence shifts and computer simulations revealed that dynamics and hydration at the tunnel mouth differ substantially between the redesigned and target dehalogenase.
authors.affiliation: 1] J. Heyrovsky Institute of Physical Chemistry of the ASCR, v.v.i., Academy of Sciences of the Czech Republic, Prague, Czech Republic. [2].
authors.forename: Jan
authors.initials: J
authors.lastname: Sykora
authors.affiliation: 1] Loschmidt Laboratories, Department of Experimental Biology and Research Centre for Toxic Compounds in the Environment (RECETOX), Faculty of Science, Masaryk University, Brno, Czech Republic. [2].
authors.forename: Jan
authors.initials: J
authors.lastname: Brezovsky
authors.affiliation: 1] Loschmidt Laboratories, Department of Experimental Biology and Research Centre for Toxic Compounds in the Environment (RECETOX), Faculty of Science, Masaryk University, Brno, Czech Republic. [2].
authors.forename: Tana
authors.identifier: 0000000305785268
authors.initials: T
authors.lastname: Koudelakova
authors.affiliation: 1] Faculty of Science, University of South Bohemia in Ceske Budejovice, Ceske Budejovice, Czech Republic. [2] Institute of Nanobiology and Structural Biology ASCR, v.v.i., Nove Hrady, Czech Republic.
authors.forename: Maryna
authors.initials: M
authors.lastname: Lahoda
authors.affiliation: Loschmidt Laboratories, Department of Experimental Biology and Research Centre for Toxic Compounds in the Environment (RECETOX), Faculty of Science, Masaryk University, Brno, Czech Republic.
authors.forename: Andrea
authors.initials: A
authors.lastname: Fortova
authors.affiliation: J. Heyrovsky Institute of Physical Chemistry of the ASCR, v.v.i., Academy of Sciences of the Czech Republic, Prague, Czech Republic.
authors.forename: Tatsiana
authors.initials: T
authors.lastname: Chernovets
authors.affiliation: Loschmidt Laboratories, Department of Experimental Biology and Research Centre for Toxic Compounds in the Environment (RECETOX), Faculty of Science, Masaryk University, Brno, Czech Republic.
authors.forename: Radka
authors.initials: R
authors.lastname: Chaloupkova
authors.affiliation: Loschmidt Laboratories, Department of Experimental Biology and Research Centre for Toxic Compounds in the Environment (RECETOX), Faculty of Science, Masaryk University, Brno, Czech Republic.
authors.forename: Veronika
authors.initials: V
authors.lastname: Stepankova
authors.affiliation: 1] Loschmidt Laboratories, Department of Experimental Biology and Research Centre for Toxic Compounds in the Environment (RECETOX), Faculty of Science, Masaryk University, Brno, Czech Republic. [2] International Clinical Research Center, St. Anne's University Hospital Brno, Brno, Czech Republic.
authors.forename: Zbynek
authors.initials: Z
authors.lastname: Prokop
authors.affiliation: 1] Faculty of Science, University of South Bohemia in Ceske Budejovice, Ceske Budejovice, Czech Republic. [2] Institute of Nanobiology and Structural Biology ASCR, v.v.i., Nove Hrady, Czech Republic.
authors.forename: Ivana Kuta
authors.initials: IK
authors.lastname: Smatanova
authors.affiliation: J. Heyrovsky Institute of Physical Chemistry of the ASCR, v.v.i., Academy of Sciences of the Czech Republic, Prague, Czech Republic.
authors.forename: Martin
authors.initials: M
authors.lastname: Hof
authors.affiliation: 1] Loschmidt Laboratories, Department of Experimental Biology and Research Centre for Toxic Compounds in the Environment (RECETOX), Faculty of Science, Masaryk University, Brno, Czech Republic. [2] International Clinical Research Center, St. Anne's University Hospital Brno, Brno, Czech Republic.
authors.forename: Jiri
authors.initials: J
authors.lastname: Damborsky
mesh_terms: D000595:Amino Acid Sequence; D002384:Catalysis; D020134:Catalytic Domain; D018360:Crystallography, X-Ray; D006842:Hydrocarbons, Brominated / Q000737:chemistry; D006867:Hydrolases / Q000737:chemistry* / Q000235:genetics; D056004:Molecular Dynamics Simulation; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D011487:Protein Conformation; D015202:Protein Engineering; D013050:Spectrometry, Fluorescence; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D013237:Stereoisomerism; D014867:Water / Q000737:chemistry
chemical_list: D006842:Hydrocarbons, Brominated; D014867:Water; D006867:Hydrolases; C046137:haloalkane dehalogenase
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