Dynamics and Hydration Explain Failed Functional Transformation in Dehalogenase Design.

SYKORA, J., Jan BREZOVSKÝ, Táňa KOUDELÁKOVÁ, M. LAHODA, Andrea FOŘTOVÁ, T. CHERNOVETS, Radka CHALOUPKOVÁ, Veronika ŠTĚPÁNKOVÁ, Zbyněk PROKOP, I. KUTA SMATANOVA, M. HOF and Jiří DAMBORSKÝ. Dynamics and Hydration Explain Failed Functional Transformation in Dehalogenase Design. Nature Chemical Biology. 2014, vol. 10, No 6, p. 428-430. ISSN 1552-4450.

Basic information

• Original name: Dynamics and Hydration Explain Failed Functional Transformation in Dehalogenase Design.
• Authors: SYKORA, J. (203 Czech Republic), Jan BREZOVSKÝ (203 Czech Republic, belonging to the institution), Táňa KOUDELÁKOVÁ (203 Czech Republic, belonging to the institution), M. LAHODA (112 Belarus), Andrea FOŘTOVÁ (203 Czech Republic, belonging to the institution), T. CHERNOVETS (112 Belarus), Radka CHALOUPKOVÁ (203 Czech Republic, belonging to the institution), Veronika ŠTĚPÁNKOVÁ (203 Czech Republic, belonging to the institution), Zbyněk PROKOP (203 Czech Republic, belonging to the institution), I. KUTA SMATANOVA (203 Czech Republic), M. HOF (276 Germany) and Jiří DAMBORSKÝ (203 Czech Republic, guarantor, belonging to the institution).
• Edition: Nature Chemical Biology, 2014, 1552-4450.

Other information

• Original language: English
• Type of outcome: Article in a journal
• Field of Study: 10600 1.6 Biological sciences
• Country of publisher: United States of America
• Confidentiality degree: is not subject to a state or trade secret
• Impact factor: Impact factor: 12.996
• RIV identification code: RIV/00216224:14310/14:00074209
• Organization unit: Faculty of Science
• UT WoS: 000336238200008
• Keywords in English: haloalkane dehalogenase
• Tags: AKR, rivok

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.

Links

• EE2.3.30.0037, research and development project: Name: Zaměstnáním nejlepších mladých vědců k rozvoji mezinárodní spolupráce
• GAP503/12/0572, research and development project: Name: Konstrukce syntetické metabolické dráhy pro degradaci důležitého environmentálního polutantu proteinovým a metabolickým inženýrstvím

Investor: Czech Science Foundation

• LO1214, research and development project: Name: Centrum pro výzkum toxických látek v prostředí (Acronym: RECETOX)

Investor: Ministry of Education, Youth and Sports of the CR