Crystallization and preliminary crystallographic analysis of the autoinhibited form of the tandem SH3 domain of p47(phox).
p47(phox) is a cytosolic component of the phagocyte NADPH oxidase, which is responsible for the production of the superoxide which kills invasive microorgamisms. A recombinant form of a histidine-tagged tandem SH3 domain of the p47(phox)-containing polybasic autoinhibited region was expressed in Escherichia coli and purified and crystallized by the sitting-drop vapour-diffusion method at 293 K using polyethylene glycol 6000 as a precipitant. Diffraction data were collected to 2.15 A resolution at 100 K using synchrotron radiation. The crystal belongs to space group P4(1)2(1)2 or P4(3)2(1)2, with unit-cell parameters a = 100.02, c = 44.94 A. The presence of one molecule per asymmetric unit gives a crystal volume per protein mass (V(M)) of 2.6 A(3) Da(-1) and a solvent content of 52% by volume.
Yuzawa
Satoru
S
Department of Structural Biology, Graduate School of Pharmaceutical Sciences, Hokkaido University, Sapporo 060-0812, Japan.
Suzuki
Nobuo N
NN
Fujioka
Yuko
Y
Ogura
Kenji
K
Sumimoto
Hideki
H
Inagaki
Fuyuhiko
F
D018360:Crystallography, X-Ray / Q000379:methods; D004926:Escherichia coli / Q000378:metabolism; D006801:Humans; D019255:NADPH Oxidases; D010750:Phosphoproteins / Q000737:chemistry*; D017434:Protein Structure, Tertiary; D011994:Recombinant Proteins / Q000737:chemistry; D014961:X-Ray Diffraction; D018909:src Homology Domains
D010750:Phosphoproteins; D011994:Recombinant Proteins; D019255:NADPH Oxidases; C060505:neutrophil cytosolic factor 1