8AII

High Resolution Crystal Structure of Enterococcus faecium Nicotinate Nucleotide Adenylyltransferase Complexed with Adenine

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	DIAMOND BEAMLINE I04
Synchrotron site	Diamond
Beamline	I04
Temperature [K]	100
Detector technology	PIXEL
Collection date	2022-03-03
Detector	DECTRIS EIGER2 XE 16M
Wavelength(s)	0.9795
Spacegroup name	I 21 21 21
Unit cell lengths	62.511, 65.644, 108.373
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	45.269 - 1.820
R-factor	0.2071
Rwork	0.206
R-free	0.23320
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	Homology structure generated by AlphaFold
Data reduction software	DIALS
Data scaling software	DIALS
Phasing software	MrBUMP
Refinement software	PHENIX (1.15.2_3472)

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	45.269	1.850
High resolution limit [Å]	1.820	1.820
Rmerge	0.116
Rmeas	0.121
Rpim	0.033
Number of reflections	19984	979
<I/σ(I)>	10.4	0.4
Completeness [%]	98.7	97.7
Redundancy	13.5	14
CC(1/2)	0.998	0.418

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, SITTING DROP	7.2	293	15 mg/mL EfNNAT in 50mM HEPES, 2mM DTT, 150mM NaCl, 0.02% NaN3 at pH 7.2 with the crystallization buffer consisting of 0.2 M Lithium sulfate monohydrate, 0.1 M BIS-TRIS (pH 5.5), 25% w/v Polyethylene glycol 3350