Expression, crystallization and preliminary X-ray analysis of the periplasmic stress sensory protein RseB from Escherichia coli.
Sensing external stress in the bacterial periplasm and signal transduction to the cytoplasm are important functions of the CpxAR, Bae and sigma(E) signalling pathways. In Escherichia coli, the sigma(E) pathway can be activated through degradation of the antisigma factor RseA by DegS and YaeL. The periplasmic protein RseB plays an important role in this pathway by exerting a direct or indirect negative effect on YaeL cleavage efficiency. RseB from E. coli, missing the periplasmic signal sequence (RseB(DeltaN)), was cloned, expressed, purified and crystallized. Crystals were obtained in two different forms belonging to space group P42(1)2 (form I) and C222(1) (form II) and diffracted to 2.8 and 2.4 A resolution, respectively. In crystal form I two copies of the protein were located in the asymmetric unit according to heavy-atom analysis, while crystal form II contained three copies.
Wollmann
Petra
P
Max Planck Institute of Biochemistry, Department of Membrane Biochemistry, Am Klopferspitz 18, D-82152 Martinsried, Germany. wollmann@biochem.mpg.de
Zeth
Kornelius
K
D003460:Crystallization; D018360:Crystallography, X-Ray / Q000379:methods; D004926:Escherichia coli / Q000737:chemistry* / Q000235:genetics; D029968:Escherichia coli Proteins / Q000096:biosynthesis* / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification; D015964:Gene Expression Regulation, Bacterial; D008565:Membrane Proteins / Q000096:biosynthesis* / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification; D019897:Periplasm / Q000737:chemistry* / Q000235:genetics; D021382:Protein Sorting Signals / Q000235:genetics
D029968:Escherichia coli Proteins; D008565:Membrane Proteins; D021382:Protein Sorting Signals; C494790:RseB protein, E coli