4BNP

3D structure of E. coli Isocitrate Dehydrogenase K100M mutant in complex with isocitrate and magnesium(II)

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	ROTATING ANODE
Source details	BRUKER AXS MICROSTAR
Temperature [K]	100
Detector technology	CCD
Collection date	2008-06-16
Detector	BRUKER AXS PROTEUM X8 PT135
Spacegroup name	P 43 21 2
Unit cell lengths	102.562, 102.562, 150.645
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	51.281 - 2.000
R-factor	0.1645
Rwork	0.163
R-free	0.19150
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	1ai2
RMSD bond length	0.008
RMSD bond angle	1.027
Data reduction software	SAINT
Data scaling software	SADABS
Phasing software	PHASER
Refinement software	PHENIX ((PHENIX.REFINE: 1.8.2_1309))

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	52.200	2.100
High resolution limit [Å]	2.000	2.000
Rmerge	0.080	0.360
Number of reflections	55032
<I/σ(I)>	14.5	3.1
Completeness [%]	100.0	100
Redundancy	5.6	4.3

Crystallization Conditions

crystal ID	method	pH	temperature	details
1		5.8		A SOLUTION OF THE LYS100MET MUTANT (20 MG/ML PROTEIN IN 0.9 MM CITRIC ACID, 3.5 MM NA2HPO4 PH 6.0, 100 MM NACL, 0.02% NAN3 AND 2 MM DTT) WAS MIXED 1:1 WITH A CRYSTALLIZATION SOLUTION (1.85 M NH4SO4, 50 MM CITRIC ACID/NA2HPO4, 0.1 M NACL AND 0.2 M DTT AT PH 5.8) AND 2 UL DROPS EQUILIBRATED AGAINST 500 UL OF THE CRYSTALLIZATION SOLUTION. TETRAGONAL BIPYRAMIDAL CRYSTALS, 300 UM X 150 UM X 75 UM, DEVELOPED WITHIN 5 DAYS.