8FO0

The structure of a crystallizable variant of E. coli pyruvate formate-lyase activating enzyme bound to a partially cleaved SAM molecule

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	ROTATING ANODE
Source details	RIGAKU MICROMAX-007
Temperature [K]	100
Detector technology	IMAGE PLATE
Collection date	2016-12-30
Detector	RIGAKU RAXIS IV++
Wavelength(s)	1.542
Spacegroup name	P 21 21 21
Unit cell lengths	46.580, 58.551, 84.484
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	34.260 - 1.690
R-factor	0.1653
Rwork	0.164
R-free	0.18800
Structure solution method	MOLECULAR REPLACEMENT
RMSD bond length	0.033
RMSD bond angle	0.861
Data reduction software	HKL-2000
Data scaling software	HKL-2000
Phasing software	PHASER
Refinement software	PHENIX (1.20.1-4487)

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	34.260	1.753
High resolution limit [Å]	1.690	1.690
Rmerge	0.065	0.683
Rmeas	0.067	0.721
Rpim	0.019	0.224
Number of reflections	26396	2527
<I/σ(I)>	23.76	1.8
Completeness [%]	99.7	97.15
Redundancy	11.7	8.5
CC(1/2)	0.999	0.898

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, HANGING DROP	8.5	300	4 uL of protein (10 mg/mL PFL-AE-CCR8 in 25 mM Tris, 500 200 mM KCl, with 3.5 mM SAM, 5.0 mM 7-mer RVSAYAV peptide, 0.13% glycerol, and 2.5 mM DTT) added to 1 uL of crystallization reservoir solution (30% PEG 3350, 100 mM Tris, pH 8.5) in hanging drop format over a reservoir containing of 50 uL of crystallization reservoir solution.