5HHT

Crystal structure of E. coli transketolase triple variant Ser385Tyr/Asp469Thr/Arg520Gln

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	MAX II BEAMLINE I911-3
Synchrotron site	MAX II
Beamline	I911-3
Temperature [K]	100
Detector technology	CCD
Collection date	2015-02-26
Detector	MARMOSAIC 225 mm CCD
Wavelength(s)	1.00
Spacegroup name	P 21 21 21
Unit cell lengths	89.944, 102.044, 133.076
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	44.379 - 1.500
R-factor	0.1274
Rwork	0.126
R-free	0.15690
Structure solution method	MOLECULAR REPLACEMENT
RMSD bond length	0.013
RMSD bond angle	1.573
Data reduction software	XDS
Data scaling software	XSCALE
Phasing software	PHENIX
Refinement software	PHENIX (1.9_1692)

Data quality characteristics

	Overall	Inner shell	Outer shell
Low resolution limit [Å]	44.380		1.600
High resolution limit [Å]	1.500	50.000	1.500
Rmerge	0.045		0.201
Rmeas	0.051		0.232
Total number of observations	797177
Number of reflections	190942		33781
<I/σ(I)>	22.73		7.45
Completeness [%]	97.7		99.7
Redundancy	4.2

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, HANGING DROP	7.9	281	The protein as apo-enzyme was concentrated to 16-20 mg/ml in 50 mM Gly-Gly buffer, pH 7.9. Afterwards 5 mM ThDP and 5 mM CaCl2 were added to the protein solution. Protein solution was mixed at 1+1 ratio with a reservoir solution containing 17-22 % (w/v) PEG 6000, 2 % glycerol, 50 mM Gly-Gly buffer, pH 7.9.