1K3T

Structure of Glycosomal Glyceraldehyde-3-Phosphate Dehydrogenase from Trypanosoma cruzi Complexed with Chalepin, a Coumarin Derivative Inhibitor

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	ROTATING ANODE
Source details	RIGAKU RU200
Temperature [K]	100
Detector technology	IMAGE PLATE
Collection date	1998-07-30
Detector	RIGAKU RAXIS IIC
Wavelength(s)	1.54
Spacegroup name	P 1 21 1
Unit cell lengths	82.080, 84.970, 105.240
Unit cell angles	90.00, 96.32, 90.00

Refinement procedure

Resolution	15.000 - 1.950
Rwork	0.201
R-free	0.28000
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	Holo-D-Glyceraldehyde-3-phosphate dehydrogenase from T. cruzi solved at 2.8 A resolution by Souza et al. FEBS Letters 424 (1998) 131-135.
RMSD bond length	0.150
RMSD bond angle	3.100 *
Data reduction software	DENZO
Data scaling software	SCALEPACK
Phasing software	AMoRE
Refinement software	REFMAC

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	44.720	2.020
High resolution limit [Å]	1.950	1.950
Rmerge	0.107	0.483
Total number of observations	195798 *
Number of reflections	81630
<I/σ(I)>	6.78	1.1
Completeness [%]	77.9	38.1
Redundancy	2.4	1.6

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	Vapor diffusion *	7.6 *	22 *	16% PEG 8000, 0.1M calcium acetate, 0.1M sodium cacodylate, pH 7.1, VAPOR DIFFUSION, HANGING DROP, temperature 295K

Crystallization Reagents in Literatures

ID	crystal ID	solution	reagent name	concentration (unit)	details
1	1	drop	protein	8 (mg/ml)
10	1	drop	acetonitrile	10 (%(v/v))
11	1	reservoir	sodium cacodylate	0.1 (M)	pH6.9-7.1
12	1	reservoir	calcium acetate	0.1 (M)
13	1	reservoir	PEG8000	16-18 (%)
2	1	drop	TEA	0.1 (M)	pH7.6
3	1	drop	NAD+	1 (mM)
4	1	drop	dithiothreitol	1 (mM)
5	1	drop	PMSF	1 (mM)
6	1	drop	EDTA	1 (mM)
7	1	drop	leupeptin	0.001 (mM)
8	1	drop	pepstatin pre-incubated	0.001 (mM)
9	1	drop	inhibitor	5 (mM)