5B8I

Crystal structure of Calcineurin A and Calcineurin B in complex with FKBP12 and FK506 from Coccidioides immitis RS

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	APS BEAMLINE 21-ID-G
Synchrotron site	APS
Beamline	21-ID-G
Temperature [K]	100
Detector technology	CCD
Collection date	2015-02-25
Detector	MARMOSAIC 300 mm CCD
Wavelength(s)	0.978560
Spacegroup name	P 21 21 2
Unit cell lengths	94.670, 154.630, 64.750
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	45.270 - 1.850
R-factor	0.147
Rwork	0.145
R-free	0.17200
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	1tco
RMSD bond length	0.020
RMSD bond angle	1.641
Data reduction software	XDS
Data scaling software	XSCALE
Phasing software	PHASER
Refinement software	PHENIX ((PHENIX.REFINE: DEV_1932))

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	50.000	1.900
High resolution limit [Å]	1.850	1.850
Rmerge	0.087	0.479
Number of reflections	81827
<I/σ(I)>	15.7	4.2
Completeness [%]	100.0	100
Redundancy	7.39

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, SITTING DROP	6.5	289	COCCIDIOIDES IMMITIS, COMPLEX NAMED COIMA.00174.A.GM11, AND COMPOSED OF CALCINEURIN A (COIMA.00174.A.GM11), CALCINEURIN B (COIMA.01011.A.GJ11), AND FKBP12 (COIMA.18272.A.GK11) PURIFIED IN THE PRESENCE OF FK506/ TACROLIMUS (PROTEIN BATCH #D00367) AT 10.81 MG/ML. PROTEIN BUFFER INCLUDES 25MM HEPES PH 8.0, 50 MM NACL, 5.0MM CACL2, 0.5MM TCEP, AND 5UM FK506. THE PROTEIN COMPLEX WAS CRYSTALLIZED AGAINST THE SPARSE MATRIX SCREEN PROPLEX V/V PROPANOL-1 AND CRYO-PROTECTED WITH 20% ETHYLENE GLYCOL; CRYSTAL TRACKING ID 261098D2 (HLC3-10), VAPOR DIFFUSION, SITTING DROP, TEMPERATURE 289K