2VXC

Structure of the Crb2-BRCT2 domain complex with phosphopeptide.

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	DIAMOND BEAMLINE I03
Synchrotron site	Diamond
Beamline	I03
Detector technology	CCD
Collection date	2007-05-31
Detector	ADSC CCD
Spacegroup name	P 21 21 2
Unit cell lengths	73.358, 140.415, 56.875
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	65.000 - 3.100
R-factor	0.1976
Rwork	0.195
R-free	0.24440
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	IN HOUSE STRUCTURE
RMSD bond length	0.007
RMSD bond angle	1.047
Data reduction software	MOSFLM
Data scaling software	SCALA
Phasing software	PHASER
Refinement software	PHENIX ((PHENIX.REFINE))

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	70.000	3.270
High resolution limit [Å]	3.100	3.100
Rmerge	0.090	0.350
Number of reflections	11193
<I/σ(I)>	11.5	3.7
Completeness [%]	99.8	100
Redundancy	3.8	4

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, HANGING DROP	7	293	CRB2-BRCT2 (3.5MG/ML IN 0.05M TRIS-HCL PH7.0, 0.6M NACL, 5MM DTT) WAS MIXED WITH AN EQUAL VOLUME OF PEPTIDE (0.05M TRIS-HCL PH7.0, 0.1M NACL, 5MM DTT), TO GIVE A 1:30 MOLAR RATIO OF PROTEIN:PEPTIDE. CRB2-BRCT2/PEPTIDE (7MG/ML IN 0.05M TRIS-HCL PH7.0, 1M NACL, 5MM DTT) WERE CRYSTALLISED BY HANGING DROP VAPOUR DIFFUSION AT 20C. PLATECLUSTERS GREW FROM 2UL PROTEIN, 2UL WELL SOLUTION (0.1M NA-CACODYLATE PH6.5, 7.0% PEG8000 AND 0.2M CA-ACETATE) AND 0.5UL 0.1M PR(III) ACETATE.