1PJ4

Crystal structure of human mitochondrial NAD(P)+-dependent malic enzyme in a pentary complex with natural substrate malate, ATP, Mn++, and allosteric activator fumarate.

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	APS BEAMLINE 32-ID
Synchrotron site	APS
Beamline	32-ID
Temperature [K]	100
Detector technology	CCD
Collection date	2000-12-27
Detector	ADSC QUANTUM 4
Wavelength(s)	0.9876
Spacegroup name	C 1 2 1
Unit cell lengths	226.050, 117.345, 111.310
Unit cell angles	90.00, 109.00, 90.00

Refinement procedure

Resolution	20.000 * - 2.300
R-factor	0.211
Rwork	0.211
R-free	0.25200
Structure solution method	MOLECULAR REPLACEMENT
RMSD bond length	0.007
RMSD bond angle	23.300 *
Data reduction software	DENZO
Data scaling software	SCALEPACK
Phasing software	COMO
Refinement software	CNS (1.1)

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	30.000	2.380
High resolution limit [Å]	2.300	2.300
Rmerge	0.086	0.210
Total number of observations	379634 *
Number of reflections	119648
Completeness [%]	97.6	94.2

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, HANGING DROP	7.4 *	4 *	100 mM MES (pH6.0), 10% PEG 20,000, 5% MPD, VAPOR DIFFUSION, HANGING DROP, temperature 277K

Crystallization Reagents in Literatures

ID	crystal ID	solution	reagent name	concentration (unit)	details
1	1	drop	protein	8 (mg/ml)
10	1	reservoir	PEG20000	8-10 (%(w/v))
11	1	reservoir	MPD	5 (%(v/v))
2	1	drop	Tris	30 (mM)	pH7.4
3	1	drop		30 (mM)
4	1	drop	dithiothreitol	2 (mM)
5	1	drop	ATP	10 (mM)
6	1	drop		5 (mM)
7	1	drop	fumarate	5 (mM)
8	1	drop	L-malate	10 (mM)
9	1	reservoir	MES	100 (mM)	pH6.0