6EFG

Pyruvate decarboxylase from Kluyveromyces lactis

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	EMBL/DESY, HAMBURG BEAMLINE X12
Synchrotron site	EMBL/DESY, HAMBURG
Beamline	X12
Temperature [K]	100
Detector technology	CCD
Collection date	2008-11-21
Detector	MARMOSAIC 225 mm CCD
Wavelength(s)	1.00001
Spacegroup name	P 1
Unit cell lengths	78.360, 78.480, 100.570
Unit cell angles	80.10, 67.72, 77.40

Refinement procedure

Resolution	38.250 - 2.040
R-factor	0.22123
Rwork	0.221
R-free	0.29030
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	2vk4
RMSD bond length	0.010
RMSD bond angle	1.422
Data reduction software	xia2 (0.5.492-g24d9851b-dials-1.8)
Data scaling software	Aimless (0.6.3)
Phasing software	MOLREP (11.6.04)
Refinement software	REFMAC (5.8.0230)

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	38.250	2.080
High resolution limit [Å]	2.040	2.040
Rmerge	0.086	0.659
Rmeas	0.109	0.837
Rpim	0.065	0.508
Number of reflections	126964	4511
<I/σ(I)>	8.6	1.7
Completeness [%]	93.2	66.8
Redundancy	2.7	2.3
CC(1/2)	0.996	0.689

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, HANGING DROP	6.25	281.15	2 mg/mL KlPDC in 2 mM MES, 18 mM citrate, pH 6.25, 4 mM thiamine diphosphate, 4 mM magnesium sulfate, 2 mM DTT, 1:1 with mother liquor (18-24% w/v PEG2000/PEG6000), ~25 days at 8 degrees C, soaked for 30 seconds in 2 uL mother liquor + 2 uL 15% w/v glycerol, 200 mM acetaldehyde prior to flash freezing.