4X9W

PLK-1 polo-box domain in complex with Bioactive Imidazolium-containing phosphopeptide macrocycle 4C

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	2013-11-30
Detector	PSI PILATUS 6M
Wavelength(s)	0.97950
Spacegroup name	P 1 21 1
Unit cell lengths	36.052, 51.525, 57.449
Unit cell angles	90.00, 100.55, 90.00

Refinement procedure

Resolution	38.065 - 1.798
R-factor	0.1987
Rwork	0.194
R-free	0.23880
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	3rq7
RMSD bond length	0.004
RMSD bond angle	0.846
Data reduction software	HKL-2000 ((phenix.refine: dev_1951))
Data scaling software	HKL-2000
Phasing software	PHASER
Refinement software	PHENIX ((phenix.refine: dev_1951))

Data quality characteristics

	Overall	Inner shell	Outer shell
Low resolution limit [Å]	100.000	100.000	1.830
High resolution limit [Å]	1.798	4.890	1.798
Rmerge	0.067	0.058	0.301
Rmeas	0.073	0.063	0.329
Rpim	0.028	0.024	0.132
Total number of observations	117363
Number of reflections	18528
<I/σ(I)>	7
Completeness [%]	95.7	97.5	80.2
Redundancy	6.3	6.6	5.4
CC(1/2)		0.996	0.965

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION	8	291	Frozen stocks of protein at 37 mg/mL in 10 mM TRIS pH8, 0.5 M NaCl, 10 mM DTT were thawed and diluted to 10 mg/ml with the same buffer. Complexes with each of three macrocycle compounds (3b, 3c and 4b) were prepared by adding 100 mM stocks of the macrocycle in DMSO directly to the diluted protein to achieve a final concentration of 1 mM. Crystals were grown by hanging drop vapor diffusion, with drops made by mixing equal volumes of protein-macrocycle complex and well solution containing 2-6% PEG-3350. Crystals were cryo-protected by quickly dipping in a solution of 37.5% ethylene glycol in well solution and frozen in liquid nitrogen.