2VK8

Crystal structure of the Saccharomyces cerevisiae pyruvate decarboxylase variant E477Q in complex with its substrate

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	ESRF BEAMLINE ID14-2
Synchrotron site	ESRF
Beamline	ID14-2
Temperature [K]	100
Detector technology	CCD
Collection date	2007-07-08
Detector	ADSC CCD
Spacegroup name	P 1 21 1
Unit cell lengths	78.980, 190.510, 84.140
Unit cell angles	90.00, 113.01, 90.00

Refinement procedure

Resolution	95.350 - 1.420
R-factor	0.181
Rwork	0.181
R-free	0.18600
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	2vk1
RMSD bond length	0.013
RMSD bond angle	1.250
Data reduction software	DENZO
Data scaling software	SCALEPACK
Phasing software	MOLREP (- AUTO MR)
Refinement software	REFMAC (5.2.0019)

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	99.000	1.440
High resolution limit [Å]	1.420	1.420
Rmerge	0.050	0.630
Number of reflections	427276
<I/σ(I)>	22.5	1.9
Completeness [%]	99.7	99.8
Redundancy	4.4

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, HANGING DROP	6.35	273.15	15MM CITRATE, 1.67MM MES, 315MM PYRUVATE, 1MM NADH, 0.001MG/ML ALCOHOL DEHYDROGENASE, 1.67MM TDP, 1.67MM MAGNESIUM SULFATE, 1.67MM DTT, 11.25% PEG2000, 11.25% PEG 6000, 1.1MG SCPDC/ML, PH 6.35, VAPOR DIFFUSION, HANGING DROP, TEMPERATURE 273.15K