1ML3

Evidences for a flip-flop catalytic mechanism of Trypanosoma cruzi glyceraldehyde-3-phosphate dehydrogenase, from its crystal structure in complex with reacted irreversible inhibitor 2-(2-phosphono-ethyl)-acrylic acid 4-nitro-phenyl ester

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	LNLS BEAMLINE D03B-MX1
Synchrotron site	LNLS
Beamline	D03B-MX1
Temperature [K]	100
Detector technology	IMAGE PLATE
Collection date	2000-10-05
Detector	MARRESEARCH
Wavelength(s)	1.5418
Spacegroup name	P 1 21 1
Unit cell lengths	81.456, 85.259, 105.153
Unit cell angles	90.00, 95.61, 90.00

Refinement procedure

Resolution	20.000 - 2.500
Rwork	0.199
R-free	0.26000 *
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	NATIVE STRUCTURE OF T.cruzi gGAPDH (Souza et al FEBES letters (1998) 424 131-135)
RMSD bond length	0.006
RMSD bond angle	1.220 *
Data reduction software	DENZO
Data scaling software	SCALEPACK
Phasing software	AMoRE
Refinement software	CNS (1.0)

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	20.000	2.560
High resolution limit [Å]	2.500	2.500
Rmerge	0.113	0.438
Total number of observations	134297 *
Number of reflections	48450
<I/σ(I)>	9.73	2.57
Completeness [%]	99.0 *	97.5 *
Redundancy	2.8 *	2.5 *

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, HANGING DROP	7.5	18 *	PEG 8000, CALCIUM ACETATE, EDTA, SODIUM AZIDE, pH 7.5, VAPOR DIFFUSION, HANGING DROP, temperature 291K

Crystallization Reagents in Literatures

ID	crystal ID	solution	reagent name	concentration (unit)	details
1	1	drop	protein	10 (mg/ml)
2	1	reservoir	sodium cacodylate	0.1 (M)	pH7.3-7.5
3	1	reservoir	cadmium acetate	0.1 (M)
4	1	reservoir	PEG8000	18 (%)
5	1	reservoir	EDTA	1.0 (mM)
6	1	reservoir	sodium azide	1.0 (mM)