Complexed and ligand-free high-resolution structures of urate oxidase (Uox) from Aspergillus flavus: a reassignment of the active-site binding mode.
High-resolution X-ray structures of the complexes of Aspergillus flavus urate oxidase (Uox) with three inhibitors, 8-azaxanthin (AZA), 9-methyl uric acid (MUA) and oxonic acid (OXC), were determined in an orthorhombic space group (I222). In addition, the ligand-free enzyme was also crystallized in a monoclinic form (P2(1)) and its structure determined. Higher accuracy in the three new enzyme-inhibitor complex structures (Uox-AZA, Uox-MUA and Uox-OXC) with respect to the previously determined structure of Uox-AZA (PDB code 1uox) leads to a reversed position of the inhibitor in the active site of the enzyme. The corrected anchoring of the substrate (uric acid) allows an improvement in the understanding of the enzymatic mechanism of urate oxidase.
LURE, Centre Universitaire Paris-Sud, Bâtiment 209D, BP 34, 91898 Orsay CEDEX, France. retailleau@lure.u-psud.fr
Pascal
P
Retailleau
Nathalie
N
Colloc'h
Denis
D
Vivarès
Françoise
F
Bonneté
Bertrand
B
Castro
Mohamed
M
El-Hajji
Jean Paul
JP
Mornon
Gérald
G
Monard
Thierry
T
Prangé
D000595:Amino Acid Sequence; D000818:Animals; D001231:Aspergillus flavus / Q000201:enzymology*; D001426:Bacterial Proteins / Q000737:chemistry; D020134:Catalytic Domain; D018360:Crystallography, X-Ray; D004791:Enzyme Inhibitors / Q000737:chemistry / Q000378:metabolism; D005656:Fungal Proteins / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000378:metabolism; D006801:Humans; D051379:Mice; D008968:Molecular Conformation; D008969:Molecular Sequence Data; D010094:Oxonic Acid / Q000737:chemistry / Q000378:metabolism; D011485:Protein Binding / Q000502:physiology; D017434:Protein Structure, Tertiary; D016415:Sequence Alignment; D014503:Urate Oxidase / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000378:metabolism; D014527:Uric Acid / Q000031:analogs & derivatives / Q000737:chemistry / Q000378:metabolism; D014970:Xanthines / Q000737:chemistry / Q000378:metabolism
D001426:Bacterial Proteins; D004791:Enzyme Inhibitors; D005656:Fungal Proteins; D014970:Xanthines; C018221:8-azaxanthine; D014527:Uric Acid; D010094:Oxonic Acid; D014503:Urate Oxidase