1BKH

MUCONATE LACTONIZING ENZYME FROM PSEUDOMONAS PUTIDA

Experimental procedure

Source type	ROTATING ANODE
Source details	RIGAKU RUH2R
Temperature [K]	277
Detector technology	IMAGE PLATE
Collection date	1994-02
Detector	RIGAKU RAXIS IIC
Spacegroup name	I 4 2 2
Unit cell lengths	136.180, 136.180, 265.110
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	5.000 - 2.100
R-factor	0.157
Rwork	0.157
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	UNPUBLISHED MODEL THAT WAS SOLVED BY MOLECULAR REPLACEMENT USING 1MLE AS A MODEL
RMSD bond length	0.010
RMSD bond angle	24.400 *
Data reduction software	PROCESS
Data scaling software	RIGAKU
Phasing software	X-PLOR
Refinement software	X-PLOR

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]		2.220
High resolution limit [Å]	2.000	2.100
Rmerge	0.067
Total number of observations	219744 *
Number of reflections	58765
<I/σ(I)>	8.5	2
Completeness [%]	70.0	36.1
Redundancy	3.7

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, SITTING DROP	7 *		MUCONATE LACTONIZING ENZYME (MLE; 5-10 MG/ML) WAS DIALYZED AGAINST 0.2 MM MNCL2, 7 MM MERCAPTOETHANOL, 50 MM TRIS (PH 7), AND CIS, CIS-MUCONATE WAS ADDED TO A FINAL CONCENTRATION OF 0.2 MM. CRYSTALS WERE GROWN AT ROOM TEMPERATURE BY SITTING-DROP VAPOR DIFFUSION AGAINST A WELL SOLUTION OF 70 MM NACL, 70 MM SODIUM ACETATE (PH 5.2), 0.25% POLYETHYLENE GLYCOL (AVERAGE MOLECULAR WEIGHT 3350). DROPS CONTAINED EQUAL VOLUMES (10 MICRO L) OF WELL SOLUTION AND MLE., vapor diffusion - sitting drop

Crystallization Reagents in Literatures

ID	crystal ID	solution	reagent name	concentration (unit)	details
1	1	drop	protein	5-10 (mg/ml)
2	1	drop		0.2 (mM)
3	1	drop	2-mercaptoethanol	7 (mM)
4	1	drop	Tris	50 (mM)
5	1	drop	cis,cis-muconate	0.2 (mM)
6	1	reservoir		70 (mM)
7	1	reservoir	sodium acetate	70 (mM)
8	1	reservoir	PEG3350	0.25 (%)