4HVT

Structure of a Post-proline cleaving enzyme from Rickettsia typhi

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	ALS BEAMLINE 5.0.1
Synchrotron site	ALS
Beamline	5.0.1
Temperature [K]	100
Detector technology	CCD
Collection date	2012-10-14
Detector	ADSC QUANTUM 315r
Wavelength(s)	0.9774
Spacegroup name	C 1 2 1
Unit cell lengths	199.590, 73.280, 55.460
Unit cell angles	90.00, 98.52, 90.00

Refinement procedure

Resolution	50.000 - 1.700
R-factor	0.1723
Rwork	0.170
R-free	0.20930
Structure solution method	SAD, MOLECULAR REPLACEMENT
Starting model (for MR)	An initial model was obtained from iodide SAD phasing using crystal 2 (P21). The initial model was then used as search model for C2 high resolution data set crystal 1.
RMSD bond length	0.015
RMSD bond angle	1.683
Data reduction software	XDS
Data scaling software	XSCALE
Phasing software	PHASER (2.5.2)
Refinement software	REFMAC

Data quality characteristics

	Overall	Inner shell	Outer shell
Low resolution limit [Å]	50.000	50.000	1.740
High resolution limit [Å]	1.700	7.600	1.700
Rmerge	0.065	0.025	0.493
Number of reflections	86126	1021	6348
<I/σ(I)>	16.86	42.91	3.42
Completeness [%]	99.0	98.7	98
Redundancy	4.8

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, SITTING DROP	8.5	290	Microlyic MCSG1 screen b10: 24% PEG 4000, 20% Glycerol, 160mM Magnesium Chloride, 80mM Tris:HCl pH 8.5; RityA17583bB2.PS01616 at 26.6mg/ml, cryo: EG; SG C2; for phasing crystal 2 was used SG P21, VAPOR DIFFUSION, SITTING DROP, temperature 290K
2	VAPOR DIFFUSION, SITTING DROP	7.5	290	Microlyic MCSG1 screen c5: 20% PEG 3350, 200mM Magnesium acetate; RityA17583bB2.PS01616 at 26.6mg/ml, for phasing a crystal was soaked in reservoir + 20% EG with 2.5M sodium iodide (final 500mM NaI) for 1 minute, this sample is from a differet space group (P21) than crystal 1, pH 7.5, VAPOR DIFFUSION, SITTING DROP, temperature 290K