5INJ

Crystal Structure of Prenyltransferase PriB Ternary Complex with L-Tryptophan and Dimethylallyl thiolodiphosphate (DMSPP)

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	APS BEAMLINE 31-ID
Synchrotron site	APS
Beamline	31-ID
Temperature [K]	100
Detector technology	CCD
Collection date	2015-08-14
Detector	RAYONIX MX225HE
Wavelength(s)	0.97931
Spacegroup name	C 1 2 1
Unit cell lengths	105.396, 83.422, 41.618
Unit cell angles	90.00, 106.74, 90.00

Refinement procedure

Resolution	28.816 - 1.400
R-factor	0.163
Rwork	0.162
R-free	0.19700
Structure solution method	SAD
Data reduction software	MOSFLM
Data scaling software	SCALA (3.2.5)
Phasing software	PHASER
Refinement software	PHENIX (1.10_2155)

Data quality characteristics

	Overall	Inner shell	Outer shell
Low resolution limit [Å]	28.820	28.820	1.480
High resolution limit [Å]	1.186	4.430	1.400
Rmerge	0.075	0.041	0.656
Rmeas	0.098
Total number of observations	229158
Number of reflections	65094
<I/σ(I)>	6.6	14.9	1.2
Completeness [%]	96.2	96	87.2
Redundancy	3.5	3.5	2.9

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, SITTING DROP	8.5	293.15	PriB was crystallized by sitting drop vapor diffusion method by 1:1 v/v mixing of 10 mg/mL PriB solution in the presence of 1 mM L-Tryptophan with reservoir solution containing 0.1 M Tris pH 8.5, 20% w/v PEG 3350, 0.1 M MgCl2. The mixed drops of 0.4-1 uL were equilibrated over a reservoir solution of 50 uL and incubated at 20oC in the dark. The ternary complex was obtained by 1:1.5 v/v soaking a solution of substrate mimic 25 mM Dimethylallyl S-thiolodiphosphate (DMSAPP), 0.05 M Tris pH 8.5, 10% w/v PEG 3350, 0.05 M MgCl2 into PriB crystals. The iodine derivative was prepared by 1:3 v/v soaking a solution of 0.5 M sodium iodide, 0.05 M Tris pH 8.5, 10% w/v PEG 3350, 0.05 M MgCl2 into PriB crystals