6YKA

Asymmetric [Fe]-hydrogenase from Methanolacinia paynteri apo and in complex with FeGP at 2.1-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)	0.97980
Spacegroup name	P 31 2 1
Unit cell lengths	79.481, 79.481, 179.337
Unit cell angles	90.00, 90.00, 120.00

Refinement procedure

Resolution	39.740 - 2.100
R-factor	0.197
Rwork	0.195
R-free	0.23500
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	4jjf
RMSD bond length	0.008
RMSD bond angle	1.010
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]	68.833	44.834	2.210
High resolution limit [Å]	2.100	6.640	2.100
Rmerge		0.018	0.711
Rmeas	0.062	0.019	0.746
Rpim	0.019	0.006	0.222
Number of reflections	39181	1361	5643
<I/σ(I)>	28.8	34.1	1.1
Completeness [%]	100.0	98.5	100
Redundancy	10.8	8.9	11.1
CC(1/2)	1.000		0.755

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 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 under yellow light and incubated under dark conditions. The crystals grew within two weeks in 30% w/v polyethylene glycol 4000, 200-mM lithium sulfate and 100-mM Tris pH 8.5 (JBScreen Wizard 3&4 HTS, Jena Bioscience).