6YK9

[Fe]-hydrogenase from Methanolacinia paynteri with bound guanylylpyridinol at 1.7-A resolution

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	ESRF BEAMLINE BM30A
Synchrotron site	ESRF
Beamline	BM30A
Temperature [K]	100
Detector technology	CCD
Collection date	2016-09-23
Detector	ADSC QUANTUM 315r
Wavelength(s)	0.97970
Spacegroup name	C 1 2 1
Unit cell lengths	160.827, 93.266, 83.642
Unit cell angles	90.00, 97.53, 90.00

Refinement procedure

Resolution	24.890 - 1.700
R-factor	0.19
Rwork	0.189
R-free	0.21600
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	4jjf
RMSD bond length	0.010
RMSD bond angle	1.080
Data reduction software	XDS
Data scaling software	SCALA (3.3.22)
Phasing software	MOLREP
Refinement software	BUSTER (2.10.3)

Data quality characteristics

	Overall	Inner shell	Outer shell
Low resolution limit [Å]	82.920	46.633	1.790
High resolution limit [Å]	1.700	5.380	1.700
Rmerge		0.053	0.517
Rmeas	0.125	0.063	0.603
Rpim	0.065	0.034	0.308
Number of reflections	133035	4251	19398
<I/σ(I)>	8	10.4	1.5
Completeness [%]	99.1	97.8	99.1
Redundancy	3.7	3.4	3.8
CC(1/2)	0.993		0.605

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, SITTING DROP	8.5	283.15	[Fe]-hydrogenase holoenzyme from M. paynteri was crystallized under 95%N2/5%H2 at 283.15 K using 96-well two-drop MRC crystallization plates (sitting drop vapor diffusion method). 0.7 ul of 25-mg/ml reconstituted holoenzyme was mixed with 0.7-ul reservoir solution (from crystallization kits) under yellow light and incubated under dark conditions. The best diffracting crystal came out within two weeks in 25% w/v polyethylene glycol 1500 and 100 mM SPG buffer pH 8.5 (JBScreen Wizard 3&4 HTS, Jena Bioscience). For cryo protection, the crystal was soaked in the crystallization solution supplemented with 10% v/v glycerol.