マシュー密度: 3.79 Å3/Da / 溶媒含有率: 62 % 解説: STRUCTURE DETERMINED USING THE SULFUR SAS SIGNAL RECORDED AT ID17, APS USING 1.74 ANGSTROM X-RAYS (SEE REMARK 3 REFINEMENT DETAILS)
結晶化
温度: 277 K / 手法: 蒸気拡散法, ハンギングドロップ法 / pH: 6 詳細: HANGING DROP VAPOR DIFFUSION USING 5 MICROLITER DROPS CONTAINING EQUAL VOLUMES OF PROTEIN CONCENTRATE (10 MG/ML) AND RESERVOIR SOLUTION: CONTAINING 1.4 M SODIUM CITRATE, PH 6.0., VAPOR ...詳細: HANGING DROP VAPOR DIFFUSION USING 5 MICROLITER DROPS CONTAINING EQUAL VOLUMES OF PROTEIN CONCENTRATE (10 MG/ML) AND RESERVOIR SOLUTION: CONTAINING 1.4 M SODIUM CITRATE, PH 6.0., VAPOR DIFFUSION, HANGING DROP, temperature 277K
構造決定の手法: SULFUR SAS / 解像度: 1.73→19.84 Å / Rfactor Rfree error: 0.004 / Data cutoff high absF: 260910.23 / Data cutoff low absF: 0 / 交差検証法: THROUGHOUT / σ(F): 0 / σ(I): 0 / 立体化学のターゲット値: Engh & Huber 詳細: THE STRUCTURE WAS DETERMINED USING SINGLE WAVELENGTH ANOMALOUS SCATTERING DATA RECORDED FOR THE NATIVE PROTEIN. DATA WERE RECORDED ON TWO OBELIN CRYSTALS TO 2.5 ANGSTROMS RESOLUTION AT ...詳細: THE STRUCTURE WAS DETERMINED USING SINGLE WAVELENGTH ANOMALOUS SCATTERING DATA RECORDED FOR THE NATIVE PROTEIN. DATA WERE RECORDED ON TWO OBELIN CRYSTALS TO 2.5 ANGSTROMS RESOLUTION AT BEAMLINE ID-17 (IMCA-CAT), ADVANCED PHOTON SOURCE (APS), ARGONNE NATIONAL LABORATORY USING A MARRESEARCH 165 MM CCD DETECTOR AND 1.74 ANGSTROM X-RAYS. A DATA COLLECTION STRATEGY (HKL2000) WAS DETERMINED FROM THE INITIAL IMAGE TO GIVE THE STARTING AND ENDING POINTS OF THE DATA COLLECTION TO INSURE A MINIMUM DATA COMPLETENESS OF 98 PERCENT. DATA WERE COLLECTED USING THE "FREIDEL FLIP" METHOD. DATA PROCESSING WAS CARRIED OUT USING HKL2000. THE PROCESSED DATA FORMING THE TWO DATA SETS WERE MERGED TO INCREASE REDUNDANCY (WU, ET AL., (2000) J. PROT. PEP. LETTS. 7, 25-32) AND SOLVE WAS USED TO DETERMINE THE SIX OF THE EIGHT SULFUR ATOM POSITIONS. PHASES WAS USED TO REFINE THE SULFUR POSITIONS AND TO LOCATE THE REMAINING SULFUR SITES BY BIJVOET DIFFERENCE FOURIER ANALYSIS. THREE ADDITIONAL SITES WERE IDENTIFIED SUGGESTING THAT ONE OF THE SULFUR SITES WAS MOST PROBABLY AN ANOMALOUS SOLVENT SUCH AS CHLORIDE. ALL ANOMALOUS SITES WERE TREATED AS SULFUR FOR THE PHASE CALCULATIONS. THE NINE SULFUR SITES WERE USED TO ESTIMATE THE INITIAL PROTEIN PHASES AT 3 ANGSTROM RESOLUTION USING SOLVENT FLATTENING (ISAS, WANG, (1985) METHODS ENZYMOL. 115, 90 112). A PRELIMINARY CHAIN TRACE WAS DONE USING THE 3.0 ANGSTROM MAP. SEQUENCE ASSIGNMENT WAS BASED ON 8 SULFUR ATOM POSITIONS. THE MODEL, 99 PERCENT COMPLETE WAS BUILT USING THE 3.0 ANGSTROM SAS MAP. RESIDUE 1 WAS NOT OBSERVED IN THE ELECTRON DENSITY AMPS AND IS PRESUMED TO BE DISORDERED. THE RESOLUTION OF THE STRUCTURE WAS THEN EXTENDED TO 1.73 ANGSTROMS RESOLUTION DURING THE FINAL STAGES OF REFINEMENT USING A MERGED DATA SET COLLECTED FROM A SINGLE CRYSTAL RECORDED AT APS (BEAMLINE ID19, WAVELENGTH 0.94 ANGSTROMS) AND AT ALS (BEAMLINE 5.0.2, WAVELENGTH 1.0 ANGSTROMS).