The First Mammalian Aldehyde Oxidase Crystal Structure

Coelho, Catarina; Mahro, Martin; Trincão, José; Carvalho, Alexandra T. P.; Ramos, María J.; Terao, Mineko; Garattini, Enrico; Leimkühler, Silke; Romão, Maria João

doi:10.1074/jbc.m112.390419

Cited by 86 publications

(77 citation statements)

References 51 publications

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“…Docking of GDC-0834 in AO Active Site. A homology model for AO was produced using the human sequence and the crystal structure of mouse AOX3 (PDB ID 3ZYV) (Coelho et al, 2012). Induced-fit docking was used to place DACA into the active site of AO.…”

Section: Methodsmentioning

confidence: 99%

A Novel Reaction Mediated by Human Aldehyde Oxidase: Amide Hydrolysis of GDC-0834

et al. 2015

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GDC-0834, a Bruton's tyrosine kinase inhibitor investigated as a potential treatment of rheumatoid arthritis, was previously reported to be extensively metabolized by amide hydrolysis such that no measurable levels of this compound were detected in human circulation after oral administration. In vitro studies in human liver cytosol determined that GDC-0834was rapidly hydrolyzed with a CL int of 0.511 ml/min per milligram of protein. Aldehyde oxidase (AO) and carboxylesterase (CES) were putatively identified as the enzymes responsible after cytosolic fractionation and mass spectrometry-proteomics analysis of the enzymatically active fractions. Results were confirmed by a series of kinetic experiments with inhibitors of AO, CES, and xanthine oxidase (XO), which implicated AO and CES, but not XO, as mediating GDC-0834 amide hydrolysis. Further supporting the interaction between GDC-0834 and AO, GDC-0834 was shown to be a potent reversible inhibitor of six known AO substrates with IC 50 values ranging from 0.86 to 1.87 mM. Additionally, in silico modeling studies suggest that GDC-0834 is capable of binding in the active site of AO with the amide bond of GDC-0834 near the molybdenum cofactor (MoCo), orientated in such a way to enable potential nucleophilic attack on the carbonyl of the amide bond by the hydroxyl of MoCo. Together, the in vitro and in silico results suggest the involvement of AO in the amide hydrolysis of GDC-0834.

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Section: Methodsmentioning

confidence: 99%

A Novel Reaction Mediated by Human Aldehyde Oxidase: Amide Hydrolysis of GDC-0834

et al. 2015

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“…to generate a protein structure followed by induced fit docking workflow using DACA as a ligand to refine residues within 5 Å of the DACA ligand. The template structure was from PDB ID 3ZYV of mouse AOX3 (Coelho et al, 2012). Sequence alignment used ClustalW, and the alignment did not need to be adjusted due to the high homology (79% homology) of the two primary sequences.…”

Section: Methodsmentioning

confidence: 99%

Evaluation of Rhesus Monkey and Guinea Pig Hepatic Cytosol Fractions as Models for Human Aldehyde Oxidase

2013

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Aldehyde oxidase (AOX) is a cytosolic enzyme expressed across a wide range of species, including guinea pig and rhesus monkey. These species are believed to be the best preclinical models for studying human AOX-mediated metabolism. We compared AOX activity in rhesus monkeys, guinea pigs, and humans using phthalazine and N-[2-(dimethylamino)ethyl]acridone-4-carboxamide (DACA) as substrates and raloxifene as an inhibitor. Michaelis-Menten kinetics was observed for phthalazine oxidation in rhesus monkey, guinea pig, and human liver cytosol, whereas substrate inhibition was seen with DACA oxidase activity in all three livers. Raloxifene inhibited phthalazine and DACA oxidase activity uncompetitively in guinea pig, whereas mixed-mode inhibition was seen in rhesus monkey.Our analysis of the primary sequence alignment of rhesus monkey, guinea pig, and human aldehyde oxidase isoform 1 (AOX1) along with homology modeling has led to the identification of several amino acid residue differences within the active site and substrate entrance channel of AOX1. We speculate that some of these residues might be responsible for the differences observed in activity. Overall, our data indicate that rhesus monkeys and guinea pigs would overestimate intrinsic clearance in humans and would be unsuitable to use as animal models. Our study also showed that AOX metabolism in species is substrate-dependent and no single animal model can be reliably used to predict every drug response in humans.

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“…The structure of mouse AOX3 was shown to be highly identical to the homodimeric butterfly-shaped XOR enzymes, consisting of two identical 150 kDa subunits, with differences in the substrate binding pocket (Coelho et al, 2012). The structure of each monomer of these enzymes includes a 20 kDa N-terminal 2x[2Fe-2S] domain, a 40 kDa central FADcontaining domain and an 85 kDa C-terminal molybdenum cofactor (Moco) domain (Garattini et al, 2003;Mahro et al, 2011).…”

Section: Introductionmentioning

confidence: 99%

“…The presence of some highly conserved residues in the catalytic centers and the high structural similarity between AOXs and XOR suggest common reaction mechanisms (Schumann et al, 2009;Coelho et al, 2012). Nevertheless, AOXs exhibit a broader substrate specificity than XOR (Pauff et al, 2009;Coelho et al, 2012).…”

Section: Introductionmentioning

confidence: 99%

The four aldehyde oxidases of Drosophila melanogaster have different gene expression patterns and enzyme substrate specificities

Marelja

Dambowsky

Bolis

et al. 2014

Journal of Experimental Biology

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In the genome of Drosophila melanogaster, four genes coding for aldehyde oxidases (AOX1-4) were identified on chromosome 3. Phylogenetic analysis showed that the AOX gene cluster evolved via independent duplication events in the vertebrate and invertebrate lineages. The functional role and the substrate specificity of the distinct Drosophila AOX enzymes is unknown. Two loss-of-function mutant alleles in this gene region, low pyridoxal oxidase (Po lpo ) and aldehyde oxidase-1 (Aldox-1 n1 ) are associated with a phenotype characterized by undetectable AOX enzymatic activity. However, the genes involved and the corresponding mutations have not yet been identified. In this study we characterized the activities, substrate specificities and expression profiles of the four AOX enzymes in D. melanogaster. We show that the Po lpo -associated phenotype is the consequence of a structural alteration of the AOX1 gene. We identified an 11-bp deletion in the Po lpo allele, resulting in a frameshift event, which removes the molybdenum cofactor domain of the encoded enzyme. Furthermore, we show that AOX2 activity is detectable only during metamorphosis and characterize a Minos-AOX2 insertion in this developmental gene that disrupts its activity. We demonstrate that the Aldox-1 n1 phenotype maps to the AOX3 gene and AOX4 activity is not detectable in our assays.

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The First Mammalian Aldehyde Oxidase Crystal Structure

Cited by 86 publications

References 51 publications

A Novel Reaction Mediated by Human Aldehyde Oxidase: Amide Hydrolysis of GDC-0834

A Novel Reaction Mediated by Human Aldehyde Oxidase: Amide Hydrolysis of GDC-0834

Evaluation of Rhesus Monkey and Guinea Pig Hepatic Cytosol Fractions as Models for Human Aldehyde Oxidase

The four aldehyde oxidases of Drosophila melanogaster have different gene expression patterns and enzyme substrate specificities

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