3INM

Crystal structure of human cytosolic NADP(+)-dependent isocitrate dehydrogenase R132H mutant in complex with NADPH, ALPHA-KETOGLUTARATE and CALCIUM(2+)

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	NSLS BEAMLINE X29A
Synchrotron site	NSLS
Beamline	X29A
Temperature [K]	100
Detector technology	CCD
Collection date	2009-06-13
Detector	ADSC QUANTUM 315r
Wavelength(s)	1.0809
Spacegroup name	C 2 2 21
Unit cell lengths	96.120, 274.690, 116.500
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	24.910 - 2.100
R-factor	0.222
Rwork	0.220
R-free	0.26200
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	1t0l
RMSD bond length	0.014
RMSD bond angle	1.105
Data reduction software	DENZO
Data scaling software	SCALEPACK
Phasing software	PHASER (2.1.4)
Refinement software	REFMAC

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	25.000	2.180
High resolution limit [Å]	2.100	2.100
Rmerge	0.080	0.305
Number of reflections	83121
<I/σ(I)>	20.3	2.4
Completeness [%]	92.4	58.8
Redundancy	8.6	2.1

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, HANGING DROP	6.5	293	Protein component: 8 mg/ml IDH, 20mM Tris-HCl pH 7.5, 100mM sodium chloride, 10mM NADPH, 10mM calcium chloride, 75mM alpha-ketoglutaric acid sodium salt. Precipitant: 100mM MES pH 6.5, 20% PEG 6000. Ratio of protein component to precipitant in initial hanging drop: 2:1., VAPOR DIFFUSION, HANGING DROP, temperature 293K