8FSI

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

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	APS BEAMLINE 24-ID-C
Synchrotron site	APS
Beamline	24-ID-C
Temperature [K]	100
Detector technology	PIXEL
Collection date	2017-02-22
Detector	DECTRIS PILATUS 6M-F
Wavelength(s)	0.97910
Spacegroup name	P 21 21 21
Unit cell lengths	46.405, 59.094, 83.545
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	41.770 - 1.460
R-factor	0.1522
Rwork	0.151
R-free	0.17300
Structure solution method	MOLECULAR REPLACEMENT
RMSD bond length	0.012
RMSD bond angle	1.198
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]	41.770	1.512
High resolution limit [Å]	1.460	1.460
Rmerge	0.052	0.300
Rmeas	0.056	0.321
Rpim	0.019	0.110
Number of reflections	39365	3661
<I/σ(I)>	19.76	3.66
Completeness [%]	96.9	91.96
Redundancy	7.6	7.7
CC(1/2)	0.999	0.959

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, HANGING DROP	9	300	4 uL of protein (10 mg/mL PFL-AE-CCR8 in 12.5 mM HEPES, 200 mM KCl, 3.5 mM SAM, 5.0 mM WT 7-mer PFL peptide, and 2.5 mM DTT) with 1 uL of crystallization reservoir solution (28% PEG 3350, 100 mM glycine, pH 9.0) in hanging drop format over 50 uL of crystallization reservoir solution