Crystallization and preliminary X-ray diffraction analysis of an oxidized state of Ohr from Xylella fastidiosa.
Xylella fastidiosa organic hydroperoxide-resistance protein (Ohr) is a dithiol-dependent peroxidase that is widely conserved in several pathogenic bacteria with high affinity for organic hydroperoxides. The protein was crystallized using the hanging-drop vapour-diffusion method in the presence of PEG 4000 as precipitant after treatment with organic peroxide (t-butyl hydroperoxide). X-ray diffraction data were collected to a maximum resolution of 1.8 A using a synchrotron-radiation source. The crystal belongs to the hexagonal space group P6(5)22, with unit-cell parameters a = b = 87.66, c = 160.28 A. The crystal structure was solved by molecular-replacement methods. The enzyme has a homodimeric quaternary structure similar to that observed for its homologue from Pseudomonas aeruginosa, but differs from the previous structure as the active-site residue Cys61 is oxidized. Structure refinement is in progress.
de Oliveira
Marcos Antonio
MA
Departamento de Biologia, Instituto de Biociências, Universidade de São Paulo, Rua do Matão 277, São Paulo SP, Brazil.
Netto
Luis Eduardo Soares
LE
Medrano
Francisco Javier
FJ
Barbosa
João Alexandre Ribeiro Gonçalves
JA
Alves
Simone Vidigal
SV
Cussiol
José Renato Rosa
JR
Guimarães
Beatriz Gomes
BG
D001426:Bacterial Proteins / Q000737:chemistry*; D001665:Binding Sites; D003460:Crystallization; D003545:Cysteine / Q000737:chemistry; D019281:Dimerization; D010100:Oxygen / Q000737:chemistry*; D011092:Polyethylene Glycols / Q000737:chemistry; D011487:Protein Conformation; D020836:Protein Structure, Quaternary; D020560:Proteobacteria / Q000378:metabolism*; D012997:Solvents / Q000737:chemistry; D014961:X-Ray Diffraction / Q000379:methods*; D020122:tert-Butylhydroperoxide / Q000737:chemistry
D001426:Bacterial Proteins; D012997:Solvents; C112257:organic hydroperoxide resistance protein, Xanthomonas campestris; D011092:Polyethylene Glycols; D020122:tert-Butylhydroperoxide; D003545:Cysteine; D010100:Oxygen