1E22

LYSYL-TRNA SYNTHETASE (LYSU) HEXAGONAL FORM complexed with lysine and the non-hydrolysable atp analogue amp-pcp

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	ELETTRA BEAMLINE 5.2R
Synchrotron site	ELETTRA
Beamline	5.2R
Temperature [K]	100
Detector technology	IMAGE PLATE
Collection date	1995-06-15
Detector	MARRESEARCH
Spacegroup name	P 61 2 2
Unit cell lengths	142.020, 142.020, 175.390
Unit cell angles	90.00, 90.00, 120.00

Refinement procedure

Resolution	12.000 - 2.430
R-factor	0.176
Rwork	0.176
R-free	0.22800
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	1e1o
RMSD bond length	0.006
RMSD bond angle	20.400 *
Data reduction software	MOSFLM
Data scaling software	CCP4
Phasing software	X-PLOR (3.1)
Refinement software	X-PLOR (3.1)

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	25.000	2.540
High resolution limit [Å]	2.430	2.430
Rmerge	0.065	0.099
Total number of observations	120728 *
Number of reflections	33781
<I/σ(I)>	8.1	6.3
Completeness [%]	84.7	65.1
Redundancy	3.6	3.1

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	Vapor diffusion, hanging drop *	7.5 *		THE PROTEIN WAS CONCENTRATED TO 12 MG/ML IN THE PRESENCE OF 5 MM LYSINE AND WAS CRYSTALLISED FROM 0.1 M PIPES PH 6.8, 0.5 M LICL, 20% PEG 4K, 17% GLYCEROL; THEN, A SMALL AMOUNT OF A SOLUTION CONTAINING AMP-PCP AND MGCL2 WAS ADDED TO THE DROP TO GET A FINAL CONCENTRATION OF ABOUT 5 MM.

Crystallization Reagents in Literatures

ID	crystal ID	solution	reagent name	concentration (unit)	details
1	1	drop	protein	12 (mg/ml)
2	1	drop	HEPES	20 (mM)
3	1	drop	lysine	5 (mM)
4	1	drop	beta-mercaptoethanol	2 (mM)
5	1	reservoir	PEG2000	20 (%)
6	1	reservoir		0.5 (M)
7	1	reservoir	PIPES	0.1 (M)