5O4M

Fresh crystals of HcgC from Methanococcus maripaludis cocrystallized with SAH and pyridinol

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	SLS BEAMLINE X10SA
Synchrotron site	SLS
Beamline	X10SA
Temperature [K]	100
Detector technology	PIXEL
Collection date	2017-02-17
Detector	DECTRIS PILATUS 6M-F
Wavelength(s)	1.00001
Spacegroup name	P 1 21 1
Unit cell lengths	71.875, 83.536, 99.987
Unit cell angles	90.00, 109.54, 90.00

Refinement procedure

Resolution	46.680 - 2.100
R-factor	0.2037
Rwork	0.202
R-free	0.23350
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	5o4h
RMSD bond length	0.010
RMSD bond angle	1.190
Data reduction software	XDS
Data scaling software	SCALA (3.3.22)
Phasing software	MOLREP (11.4.06)
Refinement software	BUSTER (2.10.1)

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	47.110	2.210
High resolution limit [Å]	2.100	2.100
Rmerge	0.054	0.544
Rpim	0.039	0.412
Number of reflections	63739	9372
<I/σ(I)>	11.5	1.7
Completeness [%]	97.9	98.8
Redundancy	2.6	2.5
CC(1/2)	0.998	0.804

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, SITTING DROP	7.5	281	The drop consists of 1 ul of enzyme solution containing ~5 mg/ml HcgC, 2 mM pyridinol (dissolved in 100% DMSO) and 2 mM SAH mixed with 1 ul of the reservoir solution : 100 mM HEPES/NaOH pH 7.5, 0.1 M NaCl, and 30% 2-Methyl-2,4-pentanediol. After only 4 days the crystals were immediately fished and frozen in liquid nitrogen for synchrotron radiation.