3H8T

Structure of Porphyromonas gingivalis heme-binding protein HmuY in complex with Heme

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	ESRF BEAMLINE ID23-1
Synchrotron site	ESRF
Beamline	ID23-1
Temperature [K]	100
Detector technology	CCD
Collection date	2008-07-18
Detector	ADSC QUANTUM 315r
Wavelength(s)	1.0000
Spacegroup name	P 42 21 2
Unit cell lengths	93.740, 93.740, 113.730
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	48.600 - 1.800
R-factor	0.16057
Rwork	0.160
R-free	0.18709
Structure solution method	SIRAS
RMSD bond length	0.012
RMSD bond angle	1.310
Data reduction software	XDS
Data scaling software	SCALA
Phasing software	SHELXDE
Refinement software	REFMAC (5.5.0046)

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	48.600	1.900
High resolution limit [Å]	1.800	1.800
Rmerge	0.101	0.669
Number of reflections	47598
<I/σ(I)>	17.6	4.2
Completeness [%]	100.0	99.9
Redundancy	12.9	13.1

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, SITTING DROP	6	293	Crystallization assays were performed following the sitting-drop vapor diffusion method. Reservoir solutions were prepared by a Tecan robot and 200-nL crystallization drops were dispensed on 96x2-well MRC plates (Wilden / Innovadyne) by a Cartesian nanodrop robot (Genomic Solutions) at the joint IBMB-CSIC/IRB/Barcelona Science Park High-Throughput Crystallography Platform (PAC). Best crystals as thin reddish prisms appeared in a Bruker steady-temperature crystal farm at 20 C using protein solution (26mg/mL in 5mM Tris/HCl, pH 8.0) and 2.4M (NH4)2SO4, 0.1M MES, pH6.0 as reservoir solution and D(+)-glucose monohydrate as additive (relative volumetric ratios 1:1:0.35). These conditions were successfully scaled up to the microliter range with Cryschem crystallization dishes (Hampton Research). VAPOR DIFFUSION, SITTING DROP, temperature 293K
1	VAPOR DIFFUSION, SITTING DROP	6	293	Crystallization assays were performed following the sitting-drop vapor diffusion method. Reservoir solutions were prepared by a Tecan robot and 200-nL crystallization drops were dispensed on 96x2-well MRC plates (Wilden / Innovadyne) by a Cartesian nanodrop robot (Genomic Solutions) at the joint IBMB-CSIC/IRB/Barcelona Science Park High-Throughput Crystallography Platform (PAC). Best crystals as thin reddish prisms appeared in a Bruker steady-temperature crystal farm at 20 C using protein solution (26mg/mL in 5mM Tris/HCl, pH 8.0) and 2.4M (NH4)2SO4, 0.1M MES, pH6.0 as reservoir solution and D(+)-glucose monohydrate as additive (relative volumetric ratios 1:1:0.35). These conditions were successfully scaled up to the microliter range with Cryschem crystallization dishes (Hampton Research). VAPOR DIFFUSION, SITTING DROP, temperature 293K