Preparation and X-ray crystallographic analysis of rubredoxin crystals from Desulfovibrio gigas to beyond ultra-high 0.68 A resolution.
Rubredoxin (D.g. Rd), a small non-heme iron-sulfur protein shown to function as a redox coupling protein from the sulfate reducing bacteria Desulfovibrio gigas, has been crystallized using the hanging-drop vapor diffusion method and macroseeding method. Rubredoxin crystals diffract to an ultra-high resolution 0.68 A using synchrotron radiation X-ray, and belong to the space group P2(1) with unit-cell parameters a=19.44 A, b=41.24 A, c=24.10 A, and beta=108.46 degrees. The data set of single-wavelength anomalous dispersion signal of iron in the native crystal was also collected for ab initio structure re-determination. Preliminary analysis indicates that there is one monomer with a [Fe-4S] cluster in each asymmetric unit. The crystal structure at this ultra-high resolution will reveal the details of its biological function. The crystal character and data collection strategy for ultra-high resolution will also be discussed.
Chen
Chun-Jung
CJ
X-ray Structural Biology Group, National Synchrotron Radiation Research Center, Hsinchu 30077, Taiwan. cjchen@nsrrc.org.tw
Liu
Ming-Yih
MY
Chen
Yi-Ting
YT
LeGall
Jean
J
D018360:Crystallography, X-Ray; D003901:Desulfovibrio / Q000378:metabolism*; D004583:Electrons; D007501:Iron / Q000737:chemistry; D008956:Models, Chemical; D009154:Mutation; D010084:Oxidation-Reduction; D012416:Rubredoxins / Q000737:chemistry*; D013455:Sulfur / Q000737:chemistry; D014965:X-Rays
D012416:Rubredoxins; D013455:Sulfur; D007501:Iron