6XHD

Structure of Prolinyl-5'-O-adenosine phosphoramidate

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	APS BEAMLINE 21-ID-G
Synchrotron site	APS
Beamline	21-ID-G
Temperature [K]	100
Detector technology	CCD
Collection date	2019-02-19
Detector	MARMOSAIC 300 mm CCD
Wavelength(s)	0.97857
Spacegroup name	C 1 2 1
Unit cell lengths	98.532, 30.689, 68.641
Unit cell angles	90.00, 90.56, 90.00

Refinement procedure

Resolution	49.310 - 1.510
R-factor	0.1711
Rwork	0.169
R-free	0.21710
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	1AFK (using one molecule)
RMSD bond length	0.006
RMSD bond angle	1.348
Data reduction software	HKL-2000
Data scaling software	HKL-2000
Phasing software	MOLREP
Refinement software	REFMAC (5.8.0258)

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	50.000	1.560
High resolution limit [Å]	1.510	1.510
Rmerge	0.062	0.588
Rpim	0.034	0.372
Number of reflections	31473	2688
<I/σ(I)>	25.76	1.38
Completeness [%]	96.9	83
Redundancy	4.2	3.1

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, HANGING DROP	5.5	291	PROTEIN WAS CRYSTALLIZED FROM 25% PEG 3350, 20 MM SODIUM CITRATE, PH 5.5 Prolinyl-5'-O-adenosine phosphoramidate soaking was achieved as follows. 1 uL of a stock solution of 100 mM ligand was added to 2uL of reservoir solution, to achieve a concentration of ~33 mM in the soaking solution. A few RNase A crystals were soaked for 45 - 90 minutes in the soaking solution