6AX1

Structure of human monoacylglycerol lipase bound to a covalent inhibitor

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	APS BEAMLINE 17-ID
Synchrotron site	APS
Beamline	17-ID
Temperature [K]	100
Detector technology	PIXEL
Collection date	2013-06-02
Detector	DECTRIS PILATUS 6M
Wavelength(s)	1.0
Spacegroup name	I 2 2 2
Unit cell lengths	86.360, 126.900, 137.510
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	93.260 - 2.260
R-factor	0.1647
Rwork	0.163
R-free	0.19840
Structure solution method	FOURIER SYNTHESIS
RMSD bond length	0.010
RMSD bond angle	1.080
Data reduction software	XDS
Data scaling software	SCALA (3.3.16)
Phasing software	BUSTER-TNT
Refinement software	BUSTER (2.11.5)

Data quality characteristics

	Overall	Inner shell	Outer shell
Low resolution limit [Å]	93.258	93.258	2.380
High resolution limit [Å]	2.260	7.150	2.260
Rmerge		0.071	0.428
Rmeas	0.118	0.086	0.529
Rpim	0.047	0.035	0.308
Total number of observations	202567	7347	9403
Number of reflections	34396	1236	4041
<I/σ(I)>	10.9	23.7	2.1
Completeness [%]	96.4	99.9	79
Redundancy	5.9	5.9	2.3

Crystallization Conditions

crystal 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.