1JKR

Testing the Water-Mediated HIN Recombinase DNA Recognition by Systematic Mutations

Experimental procedure

Experimental method	MOLECULAR REPLACEMENT WITH 1IJW HAVING THE PROPER DNA SUBSTITUTIONS AS THE STARTING MODEL.
Source type	SYNCHROTRON
Source details	CHESS BEAMLINE A1
Synchrotron site	CHESS
Beamline	A1
Temperature [K]	100
Detector technology	CCD
Collection date	1997-08-01
Spacegroup name	C 2 2 21
Unit cell lengths	86.087, 82.708, 44.865
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	24.800 - 2.280
R-factor	0.2687
Rwork	0.269
R-free	0.32450
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	1ijw
RMSD bond length	0.013
RMSD bond angle	1.700 *
Data reduction software	DENZO
Data scaling software	SCALEPACK
Phasing software	CNS
Refinement software	CNS

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	24.800	2.380
High resolution limit [Å]	2.280	2.280
Rmerge	0.089 *	0.252 *
Number of reflections	7102
<I/σ(I)>	15.68	2.49
Completeness [%]	93.2	72.1 *
Redundancy	17. *	2.6

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	Vapor diffusion, sitting drop *	8.5	4 *	HANGING DROP VAPOR DIFFUSION AT 4C, WITH INITIAL CONCENTRATION IN DROP OF 0.10 MM DNA, 0.06 MM HIN, 10 MM TRIS (PH 8.5), 10 MM CACL2, 23 MM NACL, 2.5% V/V PEG400, AND 1.56 MM NA CACODYLATE. RESERVOIR SOLUTION CONTAINS 100 MM TRIS (PH 8.5), 100 MM CACL2, 100 MM NACL, AND 25% PEG400. CONCENTRATION OF PEG400 IN RESERVOIR SOLUTION WAS INCREASED IN 5% INCREMENTS TO 35%., pH 8.50, VAPOR DIFFUSION, HANGING DROP

Crystallization Reagents

ID	crystal ID	solution ID	reagent name	concentration	details
1	1	1	DNA
10	1	2	NaCl
11	1	2	PEG 400
2	1	1	HIN
3	1	1	TRIS
4	1	1	CaCl2
5	1	1	NaCl
6	1	1	PEG 400
7	1	1	sodium cacodylate
8	1	2	Tris
9	1	2	CaCl2

Crystallization Reagents in Literatures

ID	crystal ID	solution	reagent name	concentration (unit)	details
1	1	reservoir		100 (mM)	pH8.5
2	1	reservoir	PEG400	25 (%)