6UK3

Crystal Structure of S-adenosyl-L-homocysteine hydrolase from Acanthamoeba castellanii with bound NAD and Adenosine

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	APS BEAMLINE 21-ID-F
Synchrotron site	APS
Beamline	21-ID-F
Temperature [K]	100
Detector technology	CCD
Collection date	2019-08-01
Detector	RAYONIX MX-300
Wavelength(s)	0.97872
Spacegroup name	P 1 21 1
Unit cell lengths	96.030, 112.730, 105.920
Unit cell angles	90.00, 114.72, 90.00

Refinement procedure

Resolution	50.000 - 1.400
R-factor	0.1301
Rwork	0.130
R-free	0.16110
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	5v96
RMSD bond length	0.005
RMSD bond angle	0.862
Data reduction software	XDS
Data scaling software	XSCALE
Phasing software	PHASER (2.8.3)
Refinement software	PHENIX (1.17rc1_3602)

Data quality characteristics

	Overall	Inner shell	Outer shell
Low resolution limit [Å]	50.000	50.000	1.440
High resolution limit [Å]	1.400	6.260	1.400
Rmerge	0.044	0.025	0.584
Rmeas	0.050	0.030	0.670
Number of reflections	400199	4528	29495
<I/σ(I)>	16.63	45.58	2.2
Completeness [%]	99.7	97.3	99.5
Redundancy	4.142	3.887	4.125
CC(1/2)	0.999	0.998	0.801

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, SITTING DROP	7.5	287	AccaA.00032.a.B1.PW38645 at 20.81 mg/ml, incubated with 2 mM NAD, SAH, mixed 1:1 with 12.5% (w/v) PEG-1000, 12.5% (w/v) PEG-3350, 12.5% (v/v) MPD, 0.1 M MOPS/HEPES-Na, pH=7.5, 0.02 M of sodium formate, ammonium acetate, trisodium citrate, sodium potassium L-tartrate, 0.2 M sodium oxamate. Tray: 311079g8, puck: ved0-8