6HR8

HMG-CoA reductase from Methanothermococcus thermolithotrophicus in complex with NADPH at 2.9 A resolution

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	ESRF BEAMLINE ID23-1
Synchrotron site	ESRF
Beamline	ID23-1
Temperature [K]	100
Detector technology	PIXEL
Collection date	2017-02-12
Detector	DECTRIS PILATUS 6M
Wavelength(s)	0.97662
Spacegroup name	P 4 21 2
Unit cell lengths	231.790, 231.790, 98.714
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	24.987 - 2.900
R-factor	0.2127
Rwork	0.211
R-free	0.24260
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	6hr7
RMSD bond length	0.003
RMSD bond angle	0.653
Data reduction software	XDS (3.3.22)
Data scaling software	SCALA
Phasing software	PHASER (2.6.0)
Refinement software	PHENIX ((1.10.1_2155: ???))

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	25.000	3.060
High resolution limit [Å]	2.900	2.900
Rmerge	0.095	1.285
Rmeas	0.102	1.377
Rpim	0.037	0.491
Number of reflections	59867	8592
<I/σ(I)>	13.3	1.6
Completeness [%]	99.8	100
Redundancy	7.3	7.7
CC(1/2)	0.999	0.427

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, SITTING DROP	8.5	291	HMGR at 10 mg/ml was supplemented with a final concentration of 2 mM NADPH. The sample was centrifuged for 5 minute at 13.000 RPM to remove any aggregates and dust. 0.7 ul of protein sample was spotted on a 96-well 2-drop MRC Crystallization Plates (Molecular Dimensions, Suffolk, UK) and 0.7 ul of reservoir solution was mixed. The best crystals were obtained after several month using a solution containing 40% v/v PEG 400, 100 mM Tris PH 8.5 and 200 mM Li2SO4.