0346383 - ÚMCH 2011 RIV DK eng J - Journal Article
Stsiapanava, A. - Dohnálek, Jan - Gavira, J. A. - Kutý, Michal - Koudeláková, T. - Damborský, J. - Kutá-Smatanová, Ivana
Atomic resolution studies of haloalkane dehalogenases DhaA04, DhaA14 and DhaA15 with engineered access tunnels.
Acta Crystallographica Section D-Biological Crystallography. Roč. 66, č. 9 (2010), s. 962-969. ISSN 0907-4449
R&D Projects: GA ČR GA310/09/1407; GA MŠMT(CZ) LC06010
Institutional research plan: CEZ:AV0Z40500505; CEZ:AV0Z60870520
Keywords : haloalkane dehalogenase DhaA * Rhodococcus rhodochrous * mutagenesis
Subject RIV: CE - Biochemistry
Impact factor: 6.326, year: 2010 ; AIS: 1.811, rok: 2010
DOI: https://doi.org/10.1107/S0907444910027101

Mutagenesis focused on the access tunnels of DhaA produced protein variants with significantly improved activity towards TCP. Three mutants of DhaA named DhaA04 (C176Y), DhaA14 (I135F) and DhaA15 (C176Y + I135F) were constructed in order to study the functional relevance of the enzyme tunnels The crystal structures of DhaA04, DhaA14 and DhaA15 with ligands present in the active site were solved and refined using diffraction data to 1.23, 0.95 and 1.22 Å, resolution, respectively. Structural comparisons of the wild type and the three mutants suggest that the tunnels play a key role in the processes of ligand exchange between the buried active site and the surrounding solvent.
Permanent Link: http://hdl.handle.net/11104/0005975