8V9S

Distinct Quaternary States, Intermediates, and Autoinhibition During Loading of the DnaB-Replicative Helicase by the Phage Lambda P Helicase Loader

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	2021-06-30
Detector	DECTRIS EIGER2 X 16M
Wavelength(s)	0.97918
Spacegroup name	P 31 2 1
Unit cell lengths	49.731, 49.731, 70.772
Unit cell angles	90.00, 90.00, 120.00

Refinement procedure

Resolution	43.070 - 1.860
R-factor	0.2276
Rwork	0.226
R-free	0.24360
Structure solution method	SAD
RMSD bond length	0.011
RMSD bond angle	1.256
Data reduction software	HKL-2000 (v721.2)
Data scaling software	HKL-2000 (v721.2)
Phasing software	PHENIX (1.20.1-4487-000)
Refinement software	PHENIX (1.20.1-4487-000)

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	43.070	1.980
High resolution limit [Å]	1.860	1.860
Rmeas	0.115	0.721
Rpim	0.047	0.364
Number of reflections	8852	1205
<I/σ(I)>	18.99	0.8
Completeness [%]	98.5
Redundancy	5.7
CC(1/2)	0.987	0.678

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, SITTING DROP		298.15	Crystals were prepared using the sitting drop vapor diffusion method by mixing either 0.1, or 0.2, or 0.4 uL of the protein solution and 0.2 uL of a series of commercially available crystallization screens (Qiagen). Optimized crystals for X-ray diffraction were grown at room temperature in a buffer consisting of 3 M to 4 M NaCl with 0.1 M HEPES-NaOH, pH 7 to 8 and required 7 days to form.