Mass: 18.015 Da / Num. of mol.: 745 / Source method: isolated from a natural source / Formula: H2O
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Details
Compound details
ENGINEERED RESIDUE IN CHAIN A, TYR 71 TO PHE ENGINEERED RESIDUE IN CHAIN B, TYR 71 TO PHE
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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: 2.8 Å3/Da / Density % sol: 56 % / Description: NONE
Crystal grow
Temperature: 293 K / Method: vapor diffusion, hanging drop / pH: 8 Details: CRYSTALS WERE GROWN AT 277 AND 293 K USING THE HANGING DROP VAPOR DIFFUSION METHOD. THE BEST CRYSTALS WERE OBTAINED BY MIXING 2 UL OF THE PROTEIN SOLUTION (18-20 MG/ML) CONTAINING 50 MM K- ...Details: CRYSTALS WERE GROWN AT 277 AND 293 K USING THE HANGING DROP VAPOR DIFFUSION METHOD. THE BEST CRYSTALS WERE OBTAINED BY MIXING 2 UL OF THE PROTEIN SOLUTION (18-20 MG/ML) CONTAINING 50 MM K-PHOSPHATE PH 8.0, 0.5 MM PLP, 1 MM DDT WITH AN EQUAL VOLUME OF THE RESERVOIR SOLUTION CONTAINING 50 MM TRIETHANOLAMINE BUFFER (PH 8.0), 0.5 MM PLP, 2 MM DDT, 0.4 M KCL, AND 35-38% (W/V) POLY(ETHYLENE GLYCOL) 5000 MONOMETHYL ETHER. COMPLEX OF Y71F TPL WITH 3-FLUORO-L-TYROSINE WAS PREPARED BY SOAKING CRYSTALS IN THE STABILIZING SOLUTION CONTAINING 10 MM 3- FLUORO-L-TYROSINE FOR ABOUT 30 SECONDS.
Resolution: 2.04→98.06 Å / Cor.coef. Fo:Fc: 0.965 / Cor.coef. Fo:Fc free: 0.94 / SU B: 7.459 / SU ML: 0.102 / Cross valid method: THROUGHOUT / ESU R: 0.158 / ESU R Free: 0.148 / Stereochemistry target values: MAXIMUM LIKELIHOOD Details: HYDROGENS HAVE BEEN ADDED IN THE RIDING POSITIONS. RESIDUE A 600 IS DISORDERED AND FOUND IN TWO CONFORMATIONS. A MAJOR PROPORTION OF ACTIVE SITE A IS OCCUPIED BY THE QUINONOID MOLECULE IN ...Details: HYDROGENS HAVE BEEN ADDED IN THE RIDING POSITIONS. RESIDUE A 600 IS DISORDERED AND FOUND IN TWO CONFORMATIONS. A MAJOR PROPORTION OF ACTIVE SITE A IS OCCUPIED BY THE QUINONOID MOLECULE IN THE TENSE STATE WITH OCCUPANCY ESTIMATED AT 0.67. THE RESIDUAL ELECTRON DENSITY SUGGESTS THAT A MINOR POPULATION OF SUBUNIT A IS IN THE OPEN CONFORMATION. DUE TO THE UNCONNECTED PEAKS IN THE ELECTRON DENSITY MAPS AND THE LOW OCCUPANCY, IT WAS NOT POSSIBLE TO MODEL THIS OPEN CONFORMATION. THE ELECTRON DENSITY IN THE ACTIVE SITE OF SUBUNIT A INDICATES THAT APART FROM THE TENSE QUINONOID THERE IS AN ADDITIONAL CHEMICAL SPECIES. THE RESIDUAL ELECTRON DENSITY IS BEST FITTED BY RELAXED QUINONOID MODELED WITH THE ESTIMATED OCCUPANCY OF 0.33 IN THE SAME GEOMETRY AS IN THE OPEN ACTIVE SITE B
Rfactor
Num. reflection
% reflection
Selection details
Rfree
0.20886
1047
1.5 %
RANDOM
Rwork
0.16339
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obs
0.16405
70272
95.67 %
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Solvent computation
Ion probe radii: 0.8 Å / Shrinkage radii: 0.8 Å / VDW probe radii: 1.4 Å / Solvent model: MASK