2BWQ

Crystal Structure of the RIM2 C2A-domain at 1.4 angstrom Resolution

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	APS BEAMLINE 19-ID
Synchrotron site	APS
Beamline	19-ID
Temperature [K]	100
Detector technology	CCD
Collection date	2004-08-13
Detector	CUSTOM
Spacegroup name	P 1 21 1
Unit cell lengths	25.455, 44.807, 55.756
Unit cell angles	90.00, 103.86, 90.00

Refinement procedure

Resolution	20.000 - 1.410
R-factor	0.183
Rwork	0.181
R-free	0.21300
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	1v27
RMSD bond length	0.019
RMSD bond angle	1.914
Data reduction software	HKL-2000
Data scaling software	HKL-2000
Phasing software	AMoRE
Refinement software	REFMAC (5.2.0003)

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	20.910	1.430
High resolution limit [Å]	1.410	1.410
Rmerge	0.020	0.180
Number of reflections	23283
<I/σ(I)>	52.9	5.2
Completeness [%]	98.5	82
Redundancy	4	2.8

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, HANGING DROP	4.5	293	RAT RIM2 C2A-DOMAIN (RESIDUES 722-859) DISSOLVED IN 20 MM MES (PH 6.0), 150 MM NACL AND 1 MM EDTA WAS CONCENTRATED TO 25 MG/ML AND CRYSTALLIZED IN 17.5% (W/V) PEG 4000, 0.2 M (NH4)2SO4, 0.1M SODIUM ACETATE (PH 4.5) AT 20 DEGREES C USING THE HANGING-DROP VAPOR-DIFFUSION METHOD. CRYSTALS APPEARED OVERNIGHT AND GREW TO A FINAL SIZE OF 0.05 X 0.05 X 0.1 MM WITHIN TWO DAYS. PRIOR TO DATA COLLECTION, CRYSTALS WERE TRANSFERRED INTO A SOLUTION OF 20% (W/V) PEG 4000, 0.15M NACL, 0.2 M (NH4)2SO4, 0.1M SODIUM ACETATE (PH 4.5) AND 15% (V/V) ETHYLENE GLYCOL, AND THEN FLASH-COOLED IN LIQUID PROPANE.