6TLK

Structure of methylene-tetrahydromethanopterin dehydrogenase from Methylorubrum extorquens AM1 in an open conformation containing NADP+ and methylene-H4MPT

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	2017-09-29
Detector	ADSC QUANTUM 315r
Wavelength(s)	0.97992
Spacegroup name	P 1 21 1
Unit cell lengths	76.923, 149.267, 78.559
Unit cell angles	90.00, 93.64, 90.00

Refinement procedure

Resolution	26.350 - 1.800
R-factor	0.167
Rwork	0.166
R-free	0.20100
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	1lu9
RMSD bond length	0.009
RMSD bond angle	1.210
Data reduction software	XDS
Data scaling software	SCALA
Phasing software	MOLREP
Refinement software	BUSTER (2.10.3)

Data quality characteristics

	Overall	Inner shell	Outer shell
Low resolution limit [Å]	78.401	43.316	1.900
High resolution limit [Å]	1.800	5.690	1.800
Rmerge	0.066	0.020	1.032
Rmeas	0.080	0.024	1.247
Rpim	0.044	0.013	0.690
Number of reflections	161189	4964	23624
<I/σ(I)>	12.2	27.1	0.8
Completeness [%]	98.9	94.7	99.5
Redundancy	3	3.1	3
CC(1/2)	0.990		0.543

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, SITTING DROP	7.5	281.15	The protein at 25 mg/ml was in 25 mM tris(hydroxymethyl)aminomethane (Tris)/HCl buffer pH 7.5 containing 150 mM NaCl, 5% glycerol and 2 mM dithiothreitol, supplemented with 2.5 mM methenyl-H4MPT and 2 mM NADP. The crystallization solution was 20% w/v polyethylene glycol 3350 and 200 mM magnesium formate. The crystallization drop contained 0.7 ul of protein solution and 0.7 ul crystallization solution