3UON

Structure of the human M2 muscarinic acetylcholine receptor bound to an antagonist

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	APS BEAMLINE 23-ID-D
Synchrotron site	APS
Beamline	23-ID-D
Temperature [K]	78
Detector technology	CCD
Collection date	2011-02-10
Detector	MARMOSAIC 300 mm CCD
Wavelength(s)	1.033
Spacegroup name	P 1 21 1
Unit cell lengths	78.170, 47.260, 88.120
Unit cell angles	90.00, 109.70, 90.00

Refinement procedure

Resolution	27.466 - 3.000
R-factor	0.2274
Rwork	0.225
R-free	0.27640
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	Beta2 adrenergic receptor T4 lysozyme (PDB ENTRY 2RH1)
RMSD bond length	0.002
RMSD bond angle	0.573
Data reduction software	HKL-2000
Data scaling software	HKL-2000
Phasing software	PHASER
Refinement software	PHENIX ((phenix.refine: 1.7.1_743))

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	27.500	3.050
High resolution limit [Å]	3.000	3.000
Rmerge	0.185	0.454
Number of reflections	11726
<I/σ(I)>	6.1	1.4
Completeness [%]	94.1	79.2
Redundancy	3.5	2.4

Crystallization Conditions

crystal ID	method	pH	temperature	details
1		7.5	293	25 to 35% PEG 300, 100 mM ammonium phosphate, 2% 2-Methyl-2,4-pentanediol, 100 mM HEPES, 10:1 monoolein:cholesterol lipid mix diluted 1.5:1 with protein in detergent buffer, Lipidic cubic phase, temperature 293K, pH 7.5
1		7.5	293	25 to 35% PEG 300, 100 mM ammonium phosphate, 2% 2-Methyl-2,4-pentanediol, 100 mM HEPES, 10:1 monoolein:cholesterol lipid mix diluted 1.5:1 with protein in detergent buffer, Lipidic cubic phase, temperature 293K, pH 7.5