caa a22 4500 60550783X CHVBK 20210128100634.0 cr unu---uuuuu 210128e20150301xx s 000 0 eng 10.1007/s00775-014-1196-4 doi (NATIONALLICENCE)springer-10.1007/s00775-014-1196-4 Aromatic aldehydes at the active site of aldehyde oxidoreductase from Desulfovibrio gigas [Elektronische Daten] reactivity and molecular details of the enzyme-substrate and enzyme-product interaction [Hugo Correia, Jacopo Marangon, Carlos Brondino, Jose Moura, Maria Romão, Pablo González, Teresa Santos-Silva] Desulfovibrio gigas aldehyde oxidoreductase (DgAOR) is a mononuclear molybdenum-containing enzyme from the xanthine oxidase (XO) family, a group of enzymes capable of catalyzing the oxidative hydroxylation of aldehydes and heterocyclic compounds. The kinetic studies reported in this work showed that DgAOR catalyzes the oxidative hydroxylation of aromatic aldehydes, but not heterocyclic compounds. NMR spectroscopy studies using 13C-labeled benzaldehyde confirmed that DgAOR catalyzes the conversion of aldehydes to the respective carboxylic acids. Steady-state kinetics in solution showed that high concentrations of the aromatic aldehydes produce substrate inhibition and in the case of 3-phenyl propionaldehyde a suicide substrate behavior. Hydroxyl-substituted aromatic aldehydes present none of these behaviors but the kinetic parameters are largely affected by the position of the OH group. High-resolution crystallographic structures obtained from single crystals of active-DgAOR soaked with benzaldehyde showed that the side chains of Phe425 and Tyr535 are important for the stabilization of the substrate in the active site. On the other hand, the X-ray data of DgAOR soaked with trans-cinnamaldehyde showed a cinnamic acid molecule in the substrate channel. The X-ray data of DgAOR soaked with 3-phenyl propionaldehyde showed clearly how high substrate concentrations inactivate the enzyme by binding covalently at the surface of the enzyme and blocking the substrate channel. The different reactivity of DgAOR versus aldehyde oxidase and XO towards aromatic aldehydes and N-heterocyclic compounds is explained on the basis of the present kinetic and structural data. SBIC, 2014 Aldehyde oxidoreductase nationallicence Molybdenum nationallicence X-ray crystallography nationallicence Enzyme kinetics nationallicence Suicide substrate nationallicence AOR : Aldehyde oxidoreductase nationallicence AMP : Adenosine monophosphate nationallicence AO : Aldehyde oxidase nationallicence CMP : Cytidine monophosphate nationallicence DCPIP : 2,6-Dichlorophenol-indophenol nationallicence Dg : Desulfovibrio gigas nationallicence DMSOr : Dimethyl sulfoxide reductase nationallicence FAD : Flavin adenine dinucleotide nationallicence HEPES : 4-(2-Hydroxyethyl)-1-piperazine ethanesulfonic acid nationallicence PCD : Pyranopterin cytidine dinucleotide nationallicence PMP : Pyranopterin monophosphate nationallicence rmsd : Root-mean-square deviation nationallicence SO : Sulfite oxidase nationallicence Tris-HCl : Tris(hydroxymethyl)aminomethane nationallicence U : Enzymatic unit nationallicence W-AOR : W-aldehyde oxidoreductase nationallicence XO : Xanthine oxidase nationallicence Correia Hugo UCIBIO@REQUIMTE, Faculdade de Ciências e Tecnologia, Universidade Nova de Lisboa, 2829-516, Caparica, Portugal aut Marangon Jacopo UCIBIO@REQUIMTE, Faculdade de Ciências e Tecnologia, Universidade Nova de Lisboa, 2829-516, Caparica, Portugal aut Brondino Carlos Departamento de Física, Facultad de Bioquímica y Ciencias Biológicas, Universidad Nacional del Litoral, S3000ZAA, Santa Fe, Argentina aut Moura Jose UCIBIO@REQUIMTE, Faculdade de Ciências e Tecnologia, Universidade Nova de Lisboa, 2829-516, Caparica, Portugal aut Romão Maria UCIBIO@REQUIMTE, Faculdade de Ciências e Tecnologia, Universidade Nova de Lisboa, 2829-516, Caparica, Portugal aut González Pablo Departamento de Física, Facultad de Bioquímica y Ciencias Biológicas, Universidad Nacional del Litoral, S3000ZAA, Santa Fe, Argentina aut Santos-Silva Teresa UCIBIO@REQUIMTE, Faculdade de Ciências e Tecnologia, Universidade Nova de Lisboa, 2829-516, Caparica, Portugal aut JBIC Journal of Biological Inorganic Chemistry Springer Berlin Heidelberg 20/2(2015-03-01), 219-229 0949-8257 20:2<219 2015 20 775 https://doi.org/10.1007/s00775-014-1196-4 text/html Onlinezugriff via DOI BK010053 XK010053 XK010000 Metadata rights reserved Springer special CC-BY-NC licence nationallicence 1 research-article jats NATIONALLICENCE NATIONALLICENCE NL-springer NATIONALLICENCE 856 40 https://doi.org/10.1007/s00775-014-1196-4 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Correia Hugo UCIBIO@REQUIMTE, Faculdade de Ciências e Tecnologia, Universidade Nova de Lisboa, 2829-516, Caparica, Portugal aut NATIONALLICENCE 700 1- Marangon Jacopo UCIBIO@REQUIMTE, Faculdade de Ciências e Tecnologia, Universidade Nova de Lisboa, 2829-516, Caparica, Portugal aut NATIONALLICENCE 700 1- Brondino Carlos Departamento de Física, Facultad de Bioquímica y Ciencias Biológicas, Universidad Nacional del Litoral, S3000ZAA, Santa Fe, Argentina aut NATIONALLICENCE 700 1- Moura Jose UCIBIO@REQUIMTE, Faculdade de Ciências e Tecnologia, Universidade Nova de Lisboa, 2829-516, Caparica, Portugal aut NATIONALLICENCE 700 1- Romão Maria UCIBIO@REQUIMTE, Faculdade de Ciências e Tecnologia, Universidade Nova de Lisboa, 2829-516, Caparica, Portugal aut NATIONALLICENCE 700 1- González Pablo Departamento de Física, Facultad de Bioquímica y Ciencias Biológicas, Universidad Nacional del Litoral, S3000ZAA, Santa Fe, Argentina aut NATIONALLICENCE 700 1- Santos-Silva Teresa UCIBIO@REQUIMTE, Faculdade de Ciências e Tecnologia, Universidade Nova de Lisboa, 2829-516, Caparica, Portugal aut NATIONALLICENCE 773 0- JBIC Journal of Biological Inorganic Chemistry Springer Berlin Heidelberg 20/2(2015-03-01), 219-229 0949-8257 20:2<219 2015 20 775