2V6G

Structure of Progesterone 5beta-Reductase from Digitalis Lanata in complex with NADP

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	BESSY BEAMLINE 14.1
Synchrotron site	BESSY
Beamline	14.1
Temperature [K]	100
Detector technology	CCD
Detector	MARRESEARCH
Spacegroup name	P 43 21 2
Unit cell lengths	78.590, 78.590, 178.100
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	39.340 - 2.300
R-factor	0.175
Rwork	0.171
R-free	0.21200
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	NADP FREE STRUCTURE
RMSD bond length	0.012
RMSD bond angle	1.343
Data reduction software	XDS
Data scaling software	XSCALE
Refinement software	REFMAC (5.2.0019)

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	40.000	2.400
High resolution limit [Å]	2.300	2.300
Rmerge	0.080	0.740
Number of reflections	25629
<I/σ(I)>	26	4.6
Completeness [%]	99.9	100
Redundancy	14.3	14.6

Crystallization Conditions

crystal ID	method	pH	temperature	details
1		5.8		CONTAINERLESS BATCH METHOD: 300 MICROL OF HIGH-DENSITY SILICON OIL WAS TRANSFERRED INTO A WELL OF A CELL CULTURE PLATE AND OVER-LAID WITH 500 MICROL OF REGULAR SILICON OIL. AT THE INTERFACE BETWEEN THE TWO LIQUIDS A DROPLET WAS DEPOSITED THAT WAS OBTAINED BY MIXING 0.4 MICROL OF H2O, 2.2 MICROL OF 5BETA-POR SOLUTION WITH NADP AND PROGESTERONE ADDED AND 1.8 MICROL OF A CRYSTALLIZATION SOLUTION CONSISTING OF 22.9 % MPD, 3.5 % PEG 8000, 0.05 M SODIUM ACETATE, 0.02 M CACL2, PH 5.8.