Crystal structure of Putative DNA-binding protein (YP_299413.1) from Ralstonia eutrophA JMP134 at 1.30 A resolution
Joint Center for Structural Genomics (JCSG)
Flat collimating mirror, toroid focusing mirror
GQELKGKY(MSE)KTPTGYL(MSE)VLRHGDNVLQNLEQLARDEHIPSASFVGIGF(MSE)SEATFGFYDFGRKQFDPKT YRNVE(MSE)AN(MSE)TGSIAWKEGKPSIHAHGTVTDGTFQGAGGHLLGLTVGTGSCEITVTVYPQRLDRFVDPEIQAN VLGLPQQ
GQELKGKYMKTPTGYLMVLRHGDNVLQNLEQLARDEHIPSASFVGIGFMSEATFGFYDFGRKQFDPKTYRNVEMANMTGS IAWKEGKPSIHAHGTVTDGTFQGAGGHLLGLTVGTGSCEITVTVYPQRLDRFVDPEIQANVLGLPQQ
0.2000M NaCl, 20.0000% PEG-1000, 0.1M Na,K-Phosphate pH 6.2, NANODROP, VAPOR DIFFUSION, SITTING DROP, temperature 277K
THE CONSTRUCT WAS EXPRESSED WITH A PURIFICATION TAG MGSDKIHHHHHHENLYFQG. THE TAG WAS REMOVED WITH TEV PROTEASE LEAVING ONLY A GLYCINE (0) FOLLOWED BY THE TARGET SEQUENCE. THE CLONED CONSTRUCT CONTAINS RESIDUES 28-173 OF THE FULL LENGTH PROTEIN.
1.HYDROGENS HAVE BEEN ADDED IN THE RIDING POSITIONS. 2.A MET-INHIBITION PROTOCOL WAS USED FOR SELENOMETHIONINE INCORPORATION DURING PROTEIN EXPRESSION. THE OCCUPANCY OF THE SE ATOMS IN THE MSE RESIDUES WAS REDUCED TO 0.75 TO ACCOUNT FOR THE REDUCED SCATTERING POWER DUE TO PARTIAL S-MET INCORPORATION. 3.UNKNOWN ELECTRON DENSITY IS OBSERVED NEAR RESIDUE 117, 119 AND 133 DURING REFINEMENT.
ANALYTICAL SIZE EXCLUSION CHROMATOGRAPHY WITH STATIC LIGHT SCATTERING SUPPORTS THE ASSIGNMENT OF A TRIMER AS A SIGNIFICANT OLIGOMERIZATION STATE.
YP_299413.1, Putative DNA-binding protein, Structural Genomics, Joint Center for Structural Genomics, JCSG, Protein Structure Initiative, PSI-2, Domain of unknown function (DUF296), Unknown function
QELKGKYMKTPTGYLMVLRHGDNVLQNLEQLARDEHIPSASFVGIGFMSEATFGFYDFGRKQFDPKTYRNVEMANMTGSI AWKEGKPSIHAHGTVTDGTFQGAGGHLLGLTVGTGSCEITVTVYPQRLDRFVDPEIQANVLGLPQQ