6HR7

HMG-CoA reductase from Methanothermococcus thermolithotrophicus apo form at 2.4 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 31 1 2
Unit cell lengths	83.322, 83.322, 211.212
Unit cell angles	90.00, 90.00, 120.00

Refinement procedure

Resolution	42.613 - 2.400
R-factor	0.1752
Rwork	0.173
R-free	0.21980
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	1dq8
RMSD bond length	0.007
RMSD bond angle	0.843
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]	42.613	2.530
High resolution limit [Å]	2.400	2.400
Rmerge	0.089	1.177
Rmeas	0.094	1.236
Rpim	0.029	0.375
Number of reflections	33184	4821
<I/σ(I)>	16.5	2.1
Completeness [%]	99.7	100
Redundancy	10.2	10.8
CC(1/2)	0.999	0.634

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, SITTING DROP	8.5	291	HMGR was concentrated until 10 mg/ml by using an Amicon Ultra-4 Centrifugation filter (30 kDa cut-off) (Millipore) and centrifuged for 5 minute 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.