5U29

Crystal structure of Cryptococcus neoformans H99 Acetyl-CoA Synthetase in complex with Ac-AMS

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	2016-11-13
Detector	MARMOSAIC 300 mm CCD
Wavelength(s)	0.978560
Spacegroup name	P 1
Unit cell lengths	71.280, 83.780, 101.600
Unit cell angles	110.33, 105.81, 87.75

Refinement procedure

Resolution	32.099 - 2.500
R-factor	0.1548
Rwork	0.153
R-free	0.20870
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	5k8f
RMSD bond length	0.007
RMSD bond angle	0.903
Data reduction software	XDS
Data scaling software	XSCALE
Refinement software	PHENIX ((1.11.1_2575: 000))

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	50.000	2.560
High resolution limit [Å]	2.500	2.500
Rmerge	0.073	0.588
Number of reflections	72292
<I/σ(I)>	15.11	2.64
Completeness [%]	98.5	97.8
Redundancy	3.9	4
CC(1/2)	0.998	0.805

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, SITTING DROP	6	289	ACETYL COA SYNTHETASE FROM CRYPTOCOCCUS NEOFORMANS (CRNEC.00629.A.FS11.PD00402) AT 10MG/ ML (IN 10 MM TRIS, PH = 7.5, 20 MM NACL) WAS SET UP IN SPARSE CRYSTALLIZATION TRIALS AT 16C. 0.5MM AC-AMS WAS ADDED TO THE PROTEIN SOLUTION AND INCUBATED FOR 5 MINUTES BEFORE SETTING UP TRIALS. CRYSTALS WERE PRODUCED BY SITTING DROP VAPOR DIFFUSION WITH AN EQUAL VOLUME COMBINATION OF THE PROTEIN/LIGAND IN AN OPTIMIZATION SCREEN BASED ON WIZARD 1 AND 2 SCREEN CONDITION E8 (13.18% W/V PEG8,000, 0.2M NACL, 0.1M NA/K PHOSPHATE PH6) AND CRYO-PROTECTED IN 20% ETHYLENE GLYCOL. CRYSTAL ID 284335C8, ZNI9-6, APS21-ID-G), PH 6.0, VAPOR DIFFUSION, SITTING DROP, TEMPERATURE 289K