1IEE

STRUCTURE OF TETRAGONAL HEN EGG WHITE LYSOZYME AT 0.94 A FROM CRYSTALS GROWN BY THE COUNTER-DIFFUSION METHOD

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	EMBL/DESY, HAMBURG BEAMLINE BW7B
Synchrotron site	EMBL/DESY, HAMBURG
Beamline	BW7B
Temperature [K]	110
Detector technology	IMAGE PLATE
Collection date	1998-12-10
Detector	MARRESEARCH
Wavelength(s)	0.8337
Spacegroup name	P 43 21 2
Unit cell lengths	77.061, 77.061, 37.223
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	31.000 - 0.940
R-factor	0.1226
Rwork	0.123
R-free	0.15090
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	1lsa
RMSD bond length	0.016
RMSD bond angle	0.037
Data reduction software	DENZO
Data scaling software	SCALEPACK
Phasing software	CNS
Refinement software	SHELXL-97

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	31.000	0.960
High resolution limit [Å]	0.940	0.940
Rmerge	0.052	0.329
Total number of observations	402070 *
Number of reflections	72390
<I/σ(I)>	15.5	2.6
Completeness [%]	98.9	88.6
Redundancy	5.5

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	unknown *	4.5	293	CRYSTALLISATION TOOK PLACE IN THE EUROPEAN SPACE AGENCY (ESA) ADVANCED PROTEIN CRYSTALLISATION FACILITY (APCF) ONBOARD THE NASA SPACE SHUTTLE'S STS-95 MISSION. A NEW APCF REACTOR WITH A LONG PROTEIN CHAMBER (67 MM) WAS DESIGNED TO PERFORM CRYSTALLIZATION USING THE COUNTER-DIFFUSION METHOD UNDER MICROGRAVITY CONDITIONS. THE PROTEIN CHAMBER WAS FILLED WITH A SOLUTION OF 99 MG/ML OF HEN EGG WHITE LYSOZYME (SUPPLIED BY SEIKAGAKU, BATCH NUMBER 100940, LOT E96302) DISSOLVED IN 0.05 M ACETATE BUFFER (PH 4.5). THE SALT CHAMBER CONTAINED 20% W/V NACL AND 0.05 M ACETATE BUFFER (PH 4.5). THE ROTATORY PLUG CONNECTING THE PROTEIN AND THE SALT CHAMBERS WAS FILLED WITH 10% W/V NACL, 0.5% W/V AGAROSE GEL AND 0.05 M ACETATE BUFFER. CRYSTALLISATION TOOK PLACE IN MICROGRAVITY AT 20+/-0.1 C., Counter-Diffusion in modified APCF reactors, temperature 293K
1	unknown *	4.5	293	CRYSTALLISATION TOOK PLACE IN THE EUROPEAN SPACE AGENCY (ESA) ADVANCED PROTEIN CRYSTALLISATION FACILITY (APCF) ONBOARD THE NASA SPACE SHUTTLE'S STS-95 MISSION. A NEW APCF REACTOR WITH A LONG PROTEIN CHAMBER (67 MM) WAS DESIGNED TO PERFORM CRYSTALLIZATION USING THE COUNTER-DIFFUSION METHOD UNDER MICROGRAVITY CONDITIONS. THE PROTEIN CHAMBER WAS FILLED WITH A SOLUTION OF 99 MG/ML OF HEN EGG WHITE LYSOZYME (SUPPLIED BY SEIKAGAKU, BATCH NUMBER 100940, LOT E96302) DISSOLVED IN 0.05 M ACETATE BUFFER (PH 4.5). THE SALT CHAMBER CONTAINED 20% W/V NACL AND 0.05 M ACETATE BUFFER (PH 4.5). THE ROTATORY PLUG CONNECTING THE PROTEIN AND THE SALT CHAMBERS WAS FILLED WITH 10% W/V NACL, 0.5% W/V AGAROSE GEL AND 0.05 M ACETATE BUFFER. CRYSTALLISATION TOOK PLACE IN MICROGRAVITY AT 20+/-0.1 C., Counter-Diffusion in modified APCF reactors, temperature 293K