Cloning, purification, crystallization and preliminary X-ray analysis of DOS heme domain, a new heme oxygen sensor in Escherichia coli.
The heme-containing PAS domain of the direct oxygen-sensor protein (DOS(H)), a bona fide oxygen-sensor protein, has been cloned from Escherichia coli strain K12 and successfully purified. The oxidized form of this protein was crystallized by the hanging-drop method with a PEG 8000-based precipitant. Preliminary X-ray diffraction studies of the PAS-domain crystal show that it belongs to the orthorhombic space group P2(1)2(1)2(1), with unit-cell parameters a = 46.1, b = 68.1, c = 82.6 A. A complete diffraction data set was collected to 1.9 A for MAD phasing. The electron-density map shows two molecules in an asymmetric unit and a unique six-coordination of the heme iron.
School of Molecular Biosciences, Washington State University, Pullman 99164-4660, USA.
HaJeung
H
Park
Christine
C
Suquet
Marina I
MI
Savenkova
James D
JD
Satterlee
ChulHee
C
Kang
D003001:Cloning, Molecular; D003460:Crystallization; D018360:Crystallography, X-Ray; D004926:Escherichia coli / Q000737:chemistry*; D006418:Heme / Q000737:chemistry*; D010100:Oxygen / Q000032:analysis*; D010957:Plasmids; D011487:Protein Conformation; D013056:Spectrophotometry, Ultraviolet
D006418:Heme; D010100:Oxygen