4JSD

The X-ray crystal structure of a thermophilic cellobiose binding protein bound with laminaribiose

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	ROTATING ANODE
Source details	RIGAKU MICROMAX-007 HF
Temperature [K]	293
Detector technology	IMAGE PLATE
Collection date	2012-09-30
Detector	RIGAKU RAXIS IV++
Wavelength(s)	1.5418
Spacegroup name	P 21 21 21
Unit cell lengths	56.641, 89.596, 108.245
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	47.876 - 2.050
R-factor	0.1849
Rwork	0.184
R-free	0.20330
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	2o7i
RMSD bond length	0.007
RMSD bond angle	1.756
Data reduction software	HKL-3000
Data scaling software	HKL-3000
Phasing software	PHASER
Refinement software	PHENIX ((phenix.refine: 1.8_1069))

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	50.000	2.120
High resolution limit [Å]	2.050	2.050
Rmerge	0.062	0.488
Number of reflections	34549
<I/σ(I)>	15.4	2.5
Completeness [%]	98.1	98.3
Redundancy	3.4	3.2

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, HANGING DROP	8	293	tmCBP was concentrated to 20 mg/mL and dialyzed into 10 mM Tris, 40 mM NaCl, 0.5 mM TCEP for crystallization. Laminaribiose or laminaripentaose was added to a final concentration of 1 mM prior to crystallization trials. Crystals were grown in drops containing 2 uL of the protein solution mixed with 2 uL of 0.2-0.3 M magnesium acetate or calcium acetate, 20-30% (wt/vol) PEG 3350 equilibrated against 900 uL of the same solution, pH 8.0, VAPOR DIFFUSION, HANGING DROP, temperature 293K