FRINK Linked Data Fragments server

Matches in SemOpenAlex for { <https://semopenalex.org/work/W2037092875> ?p ?o ?g. }

• W2037092875 endingPage "411" @default.
• W2037092875 startingPage "403" @default.
• W2037092875 abstract "beta-diketone-cleaving enzyme Dke1 is a homotetrameric Fe2+-dependent dioxygenase from Acinetobacter johnsonii. The Dke1protomer adopts a single-domain beta-barrel fold characteristic of the cupin superfamily of proteins and features a mononuclear non-haem Fe2+ centre where a triad of histidine residues, His-62, His-64 and His-104, co-ordinate the catalytic metal. To provide structure-function relationships for the peculiar metal site of Dke1 in relation to the more widespread 2-His-1-Glu/Asp binding site for non-haem Fe2+,we replaced each histidine residue individually with glutamate and asparagine and compared binding of Fe2+ and four non-native catalytically inactive metals with purified apo-forms of wild-type and mutant enzymes. Results from anaerobic equilibrium microdialysis (Fe2+) and fluorescence titration (Fe2+, Cu2+, Ni2+, Mn2+ and Zn2+) experiments revealed the presence of two broadly specific metal-binding sites in native Dke1 that bind Fe2+ with a dissociation constant (Kd) of 5 microM (site I) and approximately 0.3 mM (site II). Each mutation, except for the substitution of asparagine for His-104, disrupted binding of Fe2+, but not that of the other bivalent metal ions, at site I,while leaving metal binding at site II largely unaffected. Dke1 mutants harbouring glutamate substitutions were completely inactive and not functionally complemented by external Fe2+.The Fe2+ catalytic centre activity (kcat) of mutants with asparagine substitution of His-62 and His-104 was decreased 140- and 220-fold respectively, compared with the kcat value of 8.5 s(-1) for the wild-type enzyme in the reaction with pentane-2,4-dione.The H64N mutant was not catalytically competent, except in the presence of external Fe2+ (1 mM) which elicited about 1/1000 of wild-type activity. Therefore co-ordination of Fe2+ by Dke1 requires an uncharged metallocentre, and three histidine ligands are needed for the assembly of a fully functional catalytic site. Oxidative inactivation of Dke1 was shown to involve conversion of enzyme-bound Fe2+ into Fe3+, which is then released from the metal centre." @default.
• W2037092875 created "2016-06-24" @default.
• W2037092875 creator A5027609454 @default.
• W2037092875 creator A5051203357 @default.
• W2037092875 creator A5081056309 @default.
• W2037092875 date "2009-02-11" @default.
• W2037092875 modified "2023-10-14" @default.
• W2037092875 title "Biochemical characterization and mutational analysis of the mononuclear non-haem Fe2+ site in Dke1, a cupin-type dioxygenase from <i>Acinetobacter johnsonii</i>" @default.
• W2037092875 cites W1548770541 @default.
• W2037092875 cites W1597350167 @default.
• W2037092875 cites W1654652909 @default.
• W2037092875 cites W1947079771 @default.
• W2037092875 cites W1965900569 @default.
• W2037092875 cites W1973067651 @default.
• W2037092875 cites W1977734848 @default.
• W2037092875 cites W1980294522 @default.
• W2037092875 cites W1984850597 @default.
• W2037092875 cites W1988073167 @default.
• W2037092875 cites W1991969295 @default.
• W2037092875 cites W1997201299 @default.
• W2037092875 cites W1998945596 @default.
• W2037092875 cites W2004743607 @default.
• W2037092875 cites W2005578557 @default.
• W2037092875 cites W2020108475 @default.
• W2037092875 cites W2022745658 @default.
• W2037092875 cites W2023140967 @default.
• W2037092875 cites W2031532912 @default.
• W2037092875 cites W2032730006 @default.
• W2037092875 cites W2042404460 @default.
• W2037092875 cites W2048261544 @default.
• W2037092875 cites W2048487207 @default.
• W2037092875 cites W2048935114 @default.
• W2037092875 cites W2051724875 @default.
• W2037092875 cites W2052918499 @default.
• W2037092875 cites W2057222541 @default.
• W2037092875 cites W2062996884 @default.
• W2037092875 cites W2063051256 @default.
• W2037092875 cites W2066414785 @default.
• W2037092875 cites W2066596123 @default.
• W2037092875 cites W2074555449 @default.
• W2037092875 cites W2075734597 @default.
• W2037092875 cites W2077362434 @default.
• W2037092875 cites W2078336779 @default.
• W2037092875 cites W2098380659 @default.
• W2037092875 cites W2138437172 @default.
• W2037092875 cites W2151107150 @default.
• W2037092875 cites W2154500571 @default.
• W2037092875 cites W2157473097 @default.
• W2037092875 cites W2338151994 @default.
• W2037092875 cites W2418361432 @default.
• W2037092875 cites W3002548701 @default.
• W2037092875 doi "https://doi.org/10.1042/bj20081161" @default.
• W2037092875 hasPubMedId "https://pubmed.ncbi.nlm.nih.gov/18973472" @default.
• W2037092875 hasPublicationYear "2009" @default.
• W2037092875 type Work @default.
• W2037092875 sameAs 2037092875 @default.
• W2037092875 citedByCount "28" @default.
• W2037092875 countsByYear W20370928752012 @default.
• W2037092875 countsByYear W20370928752013 @default.
• W2037092875 countsByYear W20370928752014 @default.
• W2037092875 countsByYear W20370928752015 @default.
• W2037092875 countsByYear W20370928752016 @default.
• W2037092875 countsByYear W20370928752017 @default.
• W2037092875 countsByYear W20370928752018 @default.
• W2037092875 countsByYear W20370928752020 @default.
• W2037092875 countsByYear W20370928752022 @default.
• W2037092875 crossrefType "journal-article" @default.
• W2037092875 hasAuthorship W2037092875A5027609454 @default.
• W2037092875 hasAuthorship W2037092875A5051203357 @default.
• W2037092875 hasAuthorship W2037092875A5081056309 @default.
• W2037092875 hasBestOaLocation W20370928752 @default.
• W2037092875 hasConcept C104317684 @default.
• W2037092875 hasConcept C107824862 @default.
• W2037092875 hasConcept C143065580 @default.
• W2037092875 hasConcept C181199279 @default.
• W2037092875 hasConcept C185592680 @default.
• W2037092875 hasConcept C2775895851 @default.
• W2037092875 hasConcept C2776834422 @default.
• W2037092875 hasConcept C2778460671 @default.
• W2037092875 hasConcept C41183919 @default.
• W2037092875 hasConcept C55493867 @default.
• W2037092875 hasConcept C56856141 @default.
• W2037092875 hasConcept C71240020 @default.
• W2037092875 hasConcept C86803240 @default.
• W2037092875 hasConceptScore W2037092875C104317684 @default.
• W2037092875 hasConceptScore W2037092875C107824862 @default.
• W2037092875 hasConceptScore W2037092875C143065580 @default.
• W2037092875 hasConceptScore W2037092875C181199279 @default.
• W2037092875 hasConceptScore W2037092875C185592680 @default.
• W2037092875 hasConceptScore W2037092875C2775895851 @default.
• W2037092875 hasConceptScore W2037092875C2776834422 @default.
• W2037092875 hasConceptScore W2037092875C2778460671 @default.
• W2037092875 hasConceptScore W2037092875C41183919 @default.
• W2037092875 hasConceptScore W2037092875C55493867 @default.
• W2037092875 hasConceptScore W2037092875C56856141 @default.
• W2037092875 hasConceptScore W2037092875C71240020 @default.
• W2037092875 hasConceptScore W2037092875C86803240 @default.
• W2037092875 hasIssue "2" @default.

• next