1A36

TOPOISOMERASE I/DNA COMPLEX

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	SSRL BEAMLINE BL1-5
Synchrotron site	SSRL
Beamline	BL1-5
Temperature [K]	100
Detector technology	IMAGE PLATE
Collection date	1995-05
Detector	FUJI
Spacegroup name	P 1 21 1
Unit cell lengths	56.500, 118.400, 71.500
Unit cell angles	90.00, 101.20, 90.00

Refinement procedure

Resolution	8.000 - 2.800
R-factor	0.224
Rwork	0.224
R-free	0.31200
Structure solution method	RIGID BODY REFINEMENT
Starting model (for MR)	RECONSTITUTED HUMAN TOPOISOMERASE I COVALENT COMPLEX WITH 22 BASE PAIR DUPLEX DNA
RMSD bond length	0.013
RMSD bond angle	22.100 *
Data reduction software	DENZO
Data scaling software	SCALEPACK
Phasing software	X-PLOR
Refinement software	X-PLOR (3.1)

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	20.000	2.900
High resolution limit [Å]	2.800	2.800
Rmerge	0.055	0.305
Total number of observations	63314 *
Number of reflections	21217
Completeness [%]	94.7	86.8
Redundancy	3.0 *	2.5

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	Vapor diffusion, sitting drop *	7.5 *	22 *	pH 7.70

Crystallization Reagents in Literatures

ID	crystal ID	solution	reagent name	concentration (unit)	details
1	1	reservoir	Tris-HCl	2.2 (mM)	pH6.0
10	1	drop	EDTA	0.3 (mM)
11	1	drop	Tris-HCl	2.8 (mM)	pH7.5
12	1	drop	dithiothreitol	1.4 (mM)
2	1	reservoir	MES-HCl	8.6 (mM)	pH6.8
3	1	reservoir	PEG400	12 (%(v/v))
4	1	reservoir		62 (mM)
5	1	reservoir	dithiothreitol	14 (mM)
6	1	drop	water		0.002ml
7	1	drop	oligonucleotide	0.01 (mM)
8	1	drop		0.9 (mM)
9	1	drop	protein	1.4 (mg/ml)