5FUT

Human choline kinase a1 in complex with compound 4-(dimethylamino)-1-{4-[4-(4-{[4-(pyrrolidin- 1-yl)pyridinium-1-yl]methyl}phenyl)butyl]benzyl}pyridinium (compound BR25)

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	ALBA BEAMLINE XALOC
Synchrotron site	ALBA
Beamline	XALOC
Temperature [K]	100
Detector technology	PIXEL
Detector	DECTRIS PILATUS 6M
Spacegroup name	P 43 21 2
Unit cell lengths	61.364, 61.364, 219.705
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	19.701 - 1.600
R-factor	0.1815
Rwork	0.181
R-free	0.20500
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	3g15
RMSD bond length	0.007
RMSD bond angle	1.065
Data scaling software	SCALEPACK
Phasing software	MOLREP
Refinement software	PHENIX ((1.10.1-2155_743: ???))

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	20.000	1.690
High resolution limit [Å]	1.600	1.600
Rmerge	0.040	0.670
Number of reflections	56719
<I/σ(I)>	44.2	6.6
Completeness [%]	99.9	100
Redundancy	25.4	26.2

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, SITTING DROP	7.5		HSCKALPHA1 AT 20 MG/ML WAS PREINCUBATED AT ROOM TEMPERATURE WITH 12 MM OF COMPOUND BR25 IN BUFFER 25 MM TRIS/HCL, 150 MM NACL PH 7.5 (DMSO IS AT 5% FINAL CONCENTRATION IN THE MIX). THE SITTING-DROP VAPOUR DIFFUSION METHOD WAS USED TO PRODUCE CRYSTALS BY MIXING 0.5 MICROL OF THE PROTEIN SOLUTION AND AN EQUAL VOLUME OF MOTHER LIQUOR (20% PEG 5000 MME AND 0.2 M KSCN). TETRAGONAL CRYSTALS (SPACE GROUP P43212) GREW WITHIN 2-4 DAYS AND THEN WERE SOAKED FOR ONE DAY WITH 20 MM COMPOUND BR25 SOLVED IN 100% DMSO. THE CRYSTALS USED IN THIS STUDY WERE CRYOPROTECTED IN MOTHER LIQUOR SOLUTIONS CONTAINING 20 % ETHYLENGLYCOL AND FROZEN IN A NITROGEN GAS STREAM COOLED TO 100 K.