3IOM

Crystal structure of Purine Nucleoside Phosphorylase from Mycobacterium tuberculosis in complex with 2'-Deoxyguanosine

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	CCD
Collection date	2009-02-02
Detector	MAR CCD 165 mm
Wavelength(s)	1.42800
Spacegroup name	H 3
Unit cell lengths	113.480, 113.480, 85.011
Unit cell angles	90.00, 90.00, 120.00

Refinement procedure

Resolution	28.340 - 2.140
R-factor	0.1792
Rwork	0.175
R-free	0.24894
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	1g2o
RMSD bond length	0.022
RMSD bond angle	1.976
Data reduction software	MOSFLM
Data scaling software	SCALA
Phasing software	AMoRE
Refinement software	REFMAC (5.2.0019)

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	28.690	2.270
High resolution limit [Å]	2.140	2.150
Rmerge	0.086	0.138
Number of reflections	22163
<I/σ(I)>	5.9	3.5
Completeness [%]	96.9	81.8
Redundancy	6.8	6.4

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, HANGING DROP	7.6	293	The protein was cocrystallized with a 1:0.5:1 stoichiometry of 500 mM Na2SO4 (Merck) and 3 mM 2dGuo (Fluka Biochemika), respectively, by hanging-drop vapor diffusion at 18 C. MtPNP protein (1 L of 25 mg mL-1 in 50 mM tris(hydroxymethyl)aminomethane (Tris) pH 7.6) had been previously mixed with an equal volume of the reservoir solution containing 100 mM Tris pH 8.0, 25% poly(ethylene glycol) (PEG) 3350, and 25 mM MgCl2 and hanging drops (3.5 L) were equilibrated against 400 L of the reservoir solution. , VAPOR DIFFUSION, HANGING DROP, temperature 293K