6TZ7

Crystal Structure of Aspergillus fumigatus Calcineurin A, Calcineurin B, FKBP12 and FK506 (Tacrolimus)

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	CLSI BEAMLINE 08ID-1
Synchrotron site	CLSI
Beamline	08ID-1
Temperature [K]	100
Detector technology	CCD
Collection date	2014-09-06
Detector	MARMOSAIC 300 mm CCD
Wavelength(s)	0.9795
Spacegroup name	P 1 21 1
Unit cell lengths	59.240, 94.460, 69.830
Unit cell angles	90.00, 109.28, 90.00

Refinement procedure

Resolution	48.119 - 2.500
R-factor	0.2011
Rwork	0.200
R-free	0.23200
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	1tco
RMSD bond length	0.002
RMSD bond angle	0.597
Data reduction software	XDS
Data scaling software	XSCALE
Phasing software	PHASER
Refinement software	PHENIX (dev_1803)

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	50.000	2.560
High resolution limit [Å]	2.500	2.500
Rmerge	0.096	0.697
Rmeas	0.112	0.810
Number of reflections	25206	1860
<I/σ(I)>	12	2.1
Completeness [%]	99.7	99.9
Redundancy	3.8	3.9
CC(1/2)	0.996	0.728

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, SITTING DROP	7.4	298	ASPERGILLUS FUMIGATUS VCID8013 [CALCINEURIN A], VCID8015 [CALCINEURIN B] AND VCID10288 [FKBP12] IN A SEC PURIFIED COMPLEX MEDIATED BY FK506/ TACROLIMUS AT 9.8 MG/ML [PROTEIN BATCH # 1393038]. PROTEIN BUFFER INCLUDES 25MM HEPES PH 8.0, 50 MM NACL, 5.0MM CACL2, AND 0.5MM TCEP. THE PROTEIN COMPLEX WAS CRYSTALLIZED AGAINST AN OPTIMIZATION SCREEN BASED ON THE SPARSE MATRIX SCREEN PACT CONDITION E9: 0.1M HEPES/NAOH, PH7.4, 0.2M POTASSIUM/SODIUM TARTRATE, 22.27% W/V PEG 3,350 AND CRYO-PROTECTED WITH 20% ETHYLENE GLYCOL