6AX1
Structure of human monoacylglycerol lipase bound to a covalent inhibitor
X-RAY DIFFRACTION
Crystallization
Crystalization Experiments | ||||
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ID | Method | pH | Temperature | Details |
1 | VAPOR DIFFUSION, SITTING DROP | 298 | Reservoir Buffer: 0.07M sodium cacodylate pH 5.1-5.9 and 33-51% MPD The protein is 20 mg/mL in a buffer of 15 mM HEPES pH 8.2; 2 mM TCEP; and 10% glycerol, with hexaethylene glycol monododecyl ether added to 0.1 mM. Apo protein crystals obtained in this manner were transferred to a cryo-protectant solution consisting of 70 mM NaCacodylate pH 5.1; 10% MPD; 30% PEG-MME-2K, and 1mM of inhibitor compound. Crystals were soaked overnight, then flash-frozen in LN2. |
Crystal Properties | |
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Matthews coefficient | Solvent content |
2.46 | 50.04 |
Crystal Data
Unit Cell | |
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Length ( Å ) | Angle ( ˚ ) |
a = 86.36 | α = 90 |
b = 126.9 | β = 90 |
c = 137.51 | γ = 90 |
Symmetry | |
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Space Group | I 2 2 2 |
Diffraction
Diffraction Experiment | ||||||||||||||
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ID # | Crystal ID | Scattering Type | Data Collection Temperature | Detector | Detector Type | Details | Collection Date | Monochromator | Protocol | |||||
1 | 1 | x-ray | 100 | PIXEL | DECTRIS PILATUS 6M | 2013-06-02 | M | SINGLE WAVELENGTH |
Radiation Source | |||||
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ID # | Source | Type | Wavelength List | Synchrotron Site | Beamline |
1 | SYNCHROTRON | APS BEAMLINE 17-ID | 1.0 | APS | 17-ID |
Data Collection
Overall | |||||||||||||||||||
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ID # | Resolution (High) | Resolution (Low) | Percent Possible (Observed) | R Sym I (Observed) | Rrim I (All) | Rpim I (All) | Net I Over Average Sigma (I) | Redundancy | Number Reflections (All) | Number Reflections (Observed) | Observed Criterion Sigma (F) | Observed Criterion Sigma (I) | B (Isotropic) From Wilson Plot | ||||||
1 | 2.26 | 93.258 | 96.4 | 0.099 | 0.118 | 0.047 | 10.9 | 5.9 | 34396 | 34396 | 42.42 |
Highest Resolution Shell | |||||||||||||||||||
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ID # | Resolution (High) | Resolution (Low) | Percent Possible (All) | Percent Possible (Observed) | R Merge I (Observed) | R-Sym I (Observed) | Rrim I (All) | Rpim I (All) | Mean I Over Sigma (Observed) | Redundancy | Number Unique Reflections (All) | ||||||||
1 | 2.26 | 2.38 | 79 | 0.428 | 0.428 | 0.529 | 0.308 | 1.8 | 2.3 |
Refinement
Statistics | |||||||||||||||||||
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Diffraction ID | Structure Solution Method | Cross Validation method | Resolution (High) | Resolution (Low) | Number Reflections (Observed) | Number Reflections (R-Free) | Percent Reflections (Observed) | R-Factor (Observed) | R-Work | R-Free | R-Free Selection Details | Mean Isotropic B | |||||||
X-RAY DIFFRACTION | FOURIER SYNTHESIS | THROUGHOUT | 2.26 | 93.26 | 34396 | 1735 | 96.2 | 0.1647 | 0.163 | 0.1984 | RANDOM | 44.77 |
Temperature Factor Modeling | ||||||
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Anisotropic B[1][1] | Anisotropic B[1][2] | Anisotropic B[1][3] | Anisotropic B[2][2] | Anisotropic B[2][3] | Anisotropic B[3][3] | |
-1.0284 | 4.5757 | -3.5474 |
RMS Deviations | |
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Key | Refinement Restraint Deviation |
t_other_torsion | 18.02 |
t_omega_torsion | 3.04 |
t_angle_deg | 1.08 |
t_bond_d | 0.01 |
t_dihedral_angle_d | |
t_trig_c_planes | |
t_gen_planes | |
t_it | |
t_nbd | |
t_improper_torsion |
Non-Hydrogen Atoms Used in Refinement | |
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Non-Hydrogen Atoms | Number |
Protein Atoms | 4362 |
Nucleic Acid Atoms | |
Solvent Atoms | 409 |
Heterogen Atoms | 72 |
Software
Software | |
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Software Name | Purpose |
SCALA | data scaling |
BUSTER | refinement |
PDB_EXTRACT | data extraction |
XDS | data reduction |
BUSTER-TNT | phasing |