1BWI

THE 1.8 A STRUCTURE OF MICROBATCH OIL DROP GROWN TETRAGONAL HEN EGG WHITE LYSOZYME

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	ROTATING ANODE
Source details	RIGAKU RU200
Temperature [K]	293
Detector technology	IMAGE PLATE
Detector	RIGAKU RAXIS IIC
Spacegroup name	P 43 21 2
Unit cell lengths	79.230, 79.230, 37.940
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	20.000 - 1.800
R-factor	0.171
Rwork	0.171
R-free	0.20600
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	193l
RMSD bond length	0.007
RMSD bond angle	22.200 *
Data reduction software	DENZO
Data scaling software	SCALEPACK
Refinement software	X-PLOR (3.1)

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	99.000
High resolution limit [Å]	1.700	1.700 *
Rmerge	0.077	0.151 *
Total number of observations	73591 *
Number of reflections	12081
<I/σ(I)>	22.6
Completeness [%]	87.2 *	43.2 *
Redundancy	6.1

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	microbatch method *	4.7	291	A SOLUTION CONTAINING 40 MG/ML OF LYSOZYME (SIGMA L-6876, BATCH NUMBER, LOT 65H7025, IE A DIFFERENT SIGMA SAMPLE TO THE EXPERIMENTS DESCRIBED ABOVE) IN 10 MM SODIUM CITRATE BUFFER PH 4.7 WAS MIXED IN AN EPPENDORF TUBE WITH EQUAL VOLUMES OF 12 % SODIUM CHLORIDE (BDH). THE MIXTURE WAS FILTERED THROUGH A 0.22 UM FILTER AND 8 UL DROPS WERE MANUALLY DISPENSED UNDER PARAFFIN OIL USING A GILSON MICRO PIPPETTE. OVER 20 IDENTICAL DROPS WERE SET UP AND, WITHIN A WEEK, SINGLE CRYSTALS TYPICALLY MEASURING 0.5 X 0.4 X 0.3 MM WERE OBTAINED IN MOST OF THE DROPS. CRYSTALLISATION TOOK PLACE AT A TEMPERATURE OF 18 - 20 C., drops under paraffin oil, temperature 291K

Crystallization Reagents in Literatures

ID	crystal ID	solution	reagent name	concentration (unit)	details
1	1	1	lysozyme	40 (mg/ml)
2	1	1	sodium citrate	10 (mM)
3	1	2		12 (%)