2FME

Crystal structure of the mitotic kinesin eg5 (ksp) in complex with mg-adp and (r)-4-(3-hydroxyphenyl)-n,n,7,8-tetramethyl-3,4-dihydroisoquinoline-2(1h)-carboxamide

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	ROTATING ANODE
Source details	RIGAKU FR-E
Temperature [K]	100
Detector technology	IMAGE PLATE
Collection date	2005-03-09
Detector	RIGAKU
Spacegroup name	C 1 2 1
Unit cell lengths	159.800, 80.000, 69.200
Unit cell angles	90.00, 96.50, 90.00

Refinement procedure

Resolution	40.000 - 2.100
Rwork	0.280
R-free	0.30500
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	INTERNAL MODEL OF EG5/LIGAND COMPLEX
RMSD bond length	0.008
RMSD bond angle	1.500
Data reduction software	DENZO
Data scaling software	SCALEPACK
Phasing software	AMoRE
Refinement software	CNS

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	50.000	2.180
High resolution limit [Å]	2.100	2.100
Rmerge	0.057	0.229
Number of reflections	47360
<I/σ(I)>	22.9	5.7
Completeness [%]	93.7	71.7
Redundancy	3.8	3.6

Crystallization Conditions

crystal ID	method	pH	temperature	details
1		7.8	277	1:1 reservoir: 0.1 M NaHEPES pH 7.8, 0.2 M MgCl2, 23% PEG-3350; protein: 25 mg/ml (610 uM) in 25 mM PIPES pH 6.8, 200 mM NaCl, 2 mM DTT, 5 mM MgCl2, 1 mM EGTA, 2.5 mM ADP, 0.75% DMSO, uM (R)-4-(3-HYDROXYPHENYL)-N,N,7,8-TETRAMETHYL-3,4-DIHYDROISOQUINOLINE-2(1H)-CARBOXAMIDE, vapor diffusion, hanging drop, temperature 277K, pH 7.80