3QUM

Crystal structure of human prostate specific antigen (PSA) in Fab sandwich with a high affinity and a PCa selective antibody

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	ESRF BEAMLINE ID23-2
Synchrotron site	ESRF
Beamline	ID23-2
Temperature [K]	100
Detector technology	CCD
Collection date	2007-03-17
Detector	MARMOSAIC 225 mm CCD
Wavelength(s)	0.872600
Spacegroup name	P 1 21 1
Unit cell lengths	55.308, 226.883, 118.694
Unit cell angles	90.00, 96.51, 90.00

Refinement procedure

Resolution	58.965 - 3.200
R-factor	0.2032
Rwork	0.198
R-free	0.29370
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	2zch using individual PSA and Fab as independent search models.
RMSD bond length	0.011
RMSD bond angle	1.623
Data reduction software	MOSFLM
Data scaling software	SCALA
Phasing software	MOLREP
Refinement software	PHENIX ((phenix.refine))

Data quality characteristics

	Overall	Inner shell	Outer shell
Low resolution limit [Å]	81.650	81.650	3.370
High resolution limit [Å]	3.200	10.120	3.200
Rmerge	0.145	0.071	0.791
Number of reflections	47573
<I/σ(I)>	8.4	20.4	1.7
Completeness [%]	99.6	99	98.8
Redundancy	2.5	2.5	2.4

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, SITTING DROP	6.5	295	Protein solution: 20 ul 5D3D11 at 10.7 mg/ml and 10 ul PSA at 5 mg/ml + 31 ul 5D5A5 at 6.7mg/ml in PBS. against a precipitant of 10% PEG 4,000, 20% ethylene glycol, 100 mM Na,K phosphate. Drops consisted of 1 ul protein + precipitant equilibrated by vapor diffusion. Streak seeding and macro seeding were used, pH 6.5, VAPOR DIFFUSION, SITTING DROP, temperature 295K