Crystallization and preliminary X-ray crystallographic analysis of Escherichia coli RbsD, a component of the ribose-transport system with unknown biochemical function.
The Escherichia coli high-affinity ribose-transport system consists of six proteins encoded by the rbs operon (rbsD, rbsA, rbsC, rbsB, rbsK and rbsR). Of the six components, RbsD is the only one whose function is unknown. In order to gain insights into the function of RbsD by structural analysis, we overexpressed and crystallized the protein as a first step toward this goal. RbsD was overexpressed in E. coli and crystallized using the hanging-drop vapour-diffusion method at 296 K. The crystals belong to the monoclinic space group C2, with unit-cell parameters a = 285.9, b = 92.3, c = 93.3 A, beta = 105.0 degrees. The unit cell is likely to contain 64 molecules of RbsD, with a crystal volume per protein mass (V(M)) of 2.43 A(3) Da(-1) and a solvent content of about 49.3% by volume. An equilibrium centrifugation analysis demonstrated that RbsD (MW = 15 292 Da) exists as an octamer in solution, suggesting that the asymmetric unit contains two octameric assemblies of RbsD. A native data set to 2.7 A resolution was obtained from a flash-cooled crystal.
Kim
M S
MS
National Creative Research Initiative Center for Biomolecular Recognition, Department of Life Science and Division of Molecular and Life Science, Pohang University of Science and Technology, Pohang, Kyungbuk 790-784, South Korea.
Oh
H
H
Park
C
C
Oh
B H
BH
D001426:Bacterial Proteins / Q000737:chemistry* / Q000378:metabolism; D001483:Base Sequence; D001692:Biological Transport; D002352:Carrier Proteins / Q000737:chemistry* / Q000378:metabolism; D003460:Crystallization; D018360:Crystallography, X-Ray; D017931:DNA Primers; D004926:Escherichia coli / Q000737:chemistry*; D011487:Protein Conformation; D012266:Ribose / Q000378:metabolism*
D001426:Bacterial Proteins; D002352:Carrier Proteins; D017931:DNA Primers; D012266:Ribose