Crystallization and preliminary diffraction analysis of Wzi, a member of the capsule export and assembly pathway in Escherichia coli.
External polysaccharide capsules provide a physical barrier that is employed by many species of bacteria for the purposes of host evasion and persistence. Wzi is a 53 kDa outer membrane β-barrel protein that is thought to play a role in the attachment of group 1 capsular polysaccharides to the cell surface. The purification and crystallization of an Escherichia coli homologue of Wzi is reported and diffraction data from native and selenomethionine-incorporated protein crystals are presented. Crystals of C-terminally His6-tagged Wzi diffracted to 2.8 Å resolution. Data processing showed that the crystals belonged to the orthorhombic space group C222, with unit-cell parameters a=128.8, b=152.8, c=94.4 Å, α=β=γ=90°. A His-tagged selenomethionine-containing variant of Wzi has also been crystallized in the same space group and diffraction data have been recorded to 3.8 Å resolution. Data processing shows that the variant crystal has similar unit-cell parameters to the native crystal.
Biomedical Sciences Research Complex, University of St Andrews, North Haugh, St Andrews KY16 9ST, Scotland.
Simon R
SR
Bushell
Hubing
H
Lou
Gregor D
GD
Wallat
Konstantinos
K
Beis
Chris
C
Whitfield
James H
JH
Naismith
D016667:Bacterial Capsules / Q000378:metabolism*; D001425:Bacterial Outer Membrane Proteins / Q000737:chemistry*; D053898:Biosynthetic Pathways; D003460:Crystallization; D018360:Crystallography, X-Ray; D004926:Escherichia coli / Q000737:chemistry*; D029968:Escherichia coli Proteins / Q000737:chemistry*; D012645:Selenomethionine / Q000737:chemistry; D014961:X-Ray Diffraction
D001425:Bacterial Outer Membrane Proteins; D029968:Escherichia coli Proteins; C556682:Wzi protein, E coli; D012645:Selenomethionine