4JSO

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

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	56.922, 89.785, 108.321
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	48.075 - 2.070
R-factor	0.1894
Rwork	0.187
R-free	0.22090
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	4jsd
RMSD bond length	0.005
RMSD bond angle	1.150
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	2.130
High resolution limit [Å]	2.070	2.070
Rmerge	0.052	0.494
Number of reflections	32940
<I/σ(I)>	16.6	3.5
Completeness [%]	95.9	98.5
Redundancy	4.3	4.2

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. 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