Structural and functional analysis of a novel haloalkane dehalogenase with two halide-binding sites
Jazyk angličtina Země Spojené státy americké Médium print-electronic
Typ dokumentu časopisecké články, práce podpořená grantem
PubMed
25004965
DOI
10.1107/s1399004714009018
PII: S1399004714009018
Knihovny.cz E-zdroje
- Klíčová slova
- catalytic activity, enzyme stability, halide-binding site, haloalkane dehalogenase, substrate specificity,
- MeSH
- analýza hlavních komponent MeSH
- halogeny metabolismus MeSH
- hydrolasy chemie metabolismus MeSH
- kinetika MeSH
- krystalizace MeSH
- vazebná místa MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- haloalkane dehalogenase MeSH Prohlížeč
- halogeny MeSH
- hydrolasy MeSH
The crystal structure of the novel haloalkane dehalogenase DbeA from Bradyrhizobium elkanii USDA94 revealed the presence of two chloride ions buried in the protein interior. The first halide-binding site is involved in substrate binding and is present in all structurally characterized haloalkane dehalogenases. The second halide-binding site is unique to DbeA. To elucidate the role of the second halide-binding site in enzyme functionality, a two-point mutant lacking this site was constructed and characterized. These substitutions resulted in a shift in the substrate-specificity class and were accompanied by a decrease in enzyme activity, stability and the elimination of substrate inhibition. The changes in enzyme catalytic activity were attributed to deceleration of the rate-limiting hydrolytic step mediated by the lower basicity of the catalytic histidine.
Citace poskytuje Crossref.org
Structural Analysis of the Ancestral Haloalkane Dehalogenase AncLinB-DmbA
Structures of hyperstable ancestral haloalkane dehalogenases show restricted conformational dynamics
PDB
4K2A
