Mass: 18.015 Da / Num. of mol.: 40 / Source method: isolated from a natural source / Formula: H2O
Sequence details
1. IN THE TARGET SEQUENCE, CYS 92 IS REPLACED BY AN SER RESIDUE BY SITE-DIRECTED MUTAGENESIS. 2. ...1. IN THE TARGET SEQUENCE, CYS 92 IS REPLACED BY AN SER RESIDUE BY SITE-DIRECTED MUTAGENESIS. 2. THE SOLUBLE DOMAIN OF MINER1 (RESIDUES 57-135) WAS EXPRESSED WITH A PURIFICATION TAG IN A PET28A(+) (NOVAGEN) BACTERIAL EXPRESSION VECTOR CONTAINING AN N-TERMINAL HIS-TAG. THE TAG WAS REMOVED WITH THROMBIN LEAVING ONLY GSHM FOLLOWED BY THE TARGET SEQUENCE.
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Experimental details
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Experiment
Experiment
Method: X-RAY DIFFRACTION / Number of used crystals: 1
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Sample preparation
Crystal
Density Matthews: 1.94 Å3/Da / Density % sol: 36.58 %
Crystal grow
Temperature: 300 K / Method: vapor diffusion / pH: 8 Details: 100 mM Tris-HCl pH 8.0, 100 mM NaCl, 15% PEG 3000, VAPOR DIFFUSION
Method to determine structure: MAD / Resolution: 2.1→37.06 Å / Cor.coef. Fo:Fc: 0.96 / Cor.coef. Fo:Fc free: 0.943 / WRfactor Rfree: 0.235 / WRfactor Rwork: 0.199 / Occupancy max: 1 / Occupancy min: 0.5 / FOM work R set: 0.928 / SU B: 7.676 / SU ML: 0.101 / SU R Cruickshank DPI: 0.208 / SU Rfree: 0.172 / TLS residual ADP flag: LIKELY RESIDUAL / Cross valid method: THROUGHOUT / σ(F): 0 / ESU R: 0.183 / ESU R Free: 0.164 Stereochemistry target values: MAXIMUM LIKELIHOOD WITH PHASES Details: 1. HYDROGENS HAVE BEEN ADDED IN THE RIDING POSITIONS. 2. ATOM RECORD CONTAINS RESIDUAL B FACTORS ONLY. 3. AN 2FE-2S CLUSTER (FES) WAS MODELED INTO EACH SUBUNIT IN THE ASYMMETRIC UNIT. THE ...Details: 1. HYDROGENS HAVE BEEN ADDED IN THE RIDING POSITIONS. 2. ATOM RECORD CONTAINS RESIDUAL B FACTORS ONLY. 3. AN 2FE-2S CLUSTER (FES) WAS MODELED INTO EACH SUBUNIT IN THE ASYMMETRIC UNIT. THE PRESENCE OF THE 2FE-2S CLUSTER WAS CORROBORATED BY ANOMALOUS DIFFERENCE MAPS. THE PROTEIN LIGANDS TO THE FE ATOMS IN THE 2FE-2S CLUSTERS ARE CYS 99, CYS 101, CYS 110, AND HIS 114.
Rfactor
Num. reflection
% reflection
Selection details
Rfree
0.216
450
5 %
RANDOM
Rwork
0.17
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obs
0.172
9013
99.23 %
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Solvent computation
Ion probe radii: 0.8 Å / Shrinkage radii: 0.8 Å / VDW probe radii: 1.2 Å / Solvent model: MASK
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