High pressure macromolecular crystallography: the 140-MPa crystal structure at 2.3 A resolution of urate oxidase, a 135-kDa tetrameric assembly.
We report the three-dimensional structure determined by high-pressure macromolecular crystallography (HPMX) of a 135-kDa homo-tetrameric enzyme, urate oxidase from Aspergillus flavus complexed with its potent inhibitor 8-azaxanthin. Urate oxidase crystals are quite sensitive to pressure, as three-dimensional order is lost at about 180 MPa. A highly complete 2.3 A resolution data set was collected at 140 MPa, close to the critical pressure. Crystal structures at atmospheric pressure and at high pressure were refined in the orthorhombic space group I222 with final crystallographic R factors 14.1% and 16.1%, respectively. The effect of pressure on temperature factors, ordered water molecules, hydrogen bond lengths, contacts, buried surface areas as well as cavity volume was investigated. Results suggest that the onset of disruption of the tetrameric assembly by pressure has been captured in the crystalline state.
Centre CYCERON, UMR 6185 Université de Caen-CNRS, Bd Becquerel, 14074 Caen, France.
Nathalie
N
Colloc'h
Eric
E
Girard
Anne-Claire
AC
Dhaussy
Richard
R
Kahn
Isabella
I
Ascone
Mohamed
M
Mezouar
Roger
R
Fourme
D001231:Aspergillus flavus / Q000201:enzymology*; D001274:Atmospheric Pressure; D018360:Crystallography, X-Ray; D004791:Enzyme Inhibitors / Q000737:chemistry*; D005656:Fungal Proteins / Q000737:chemistry*; D006874:Hydrostatic Pressure; D011487:Protein Conformation; D013696:Temperature; D014503:Urate Oxidase / Q000037:antagonists & inhibitors / Q000737:chemistry*; D014867:Water / Q000737:chemistry; D014970:Xanthines / Q000737:chemistry*
D004791:Enzyme Inhibitors; D005656:Fungal Proteins; D014970:Xanthines; D014867:Water; C018221:8-azaxanthine; D014503:Urate Oxidase