2F54

Directed evolution of human T cell receptor CDR2 residues by phage display dramatically enhances affinity for cognate peptide-MHC without increasing apparent cross-reactivity

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	SRS BEAMLINE PX14.2
Synchrotron site	SRS
Beamline	PX14.2
Temperature [K]	100
Detector technology	CCD
Collection date	2002-11-28
Detector	ADSC QUANTUM 4
Wavelength(s)	0.978
Spacegroup name	P 1 21 1
Unit cell lengths	120.020, 53.592, 152.831
Unit cell angles	90.00, 96.04, 90.00

Refinement procedure

Resolution	50.660 - 2.700
R-factor	0.225
Rwork	0.221
R-free	0.28600
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	2bnr
RMSD bond length	0.017
RMSD bond angle	1.337
Data reduction software	MOSFLM
Data scaling software	SCALA
Phasing software	AMoRE
Refinement software	REFMAC

Data quality characteristics

	Overall	Inner shell	Outer shell
Low resolution limit [Å]	50.660	50.660	2.850
High resolution limit [Å]	2.700	8.540	2.700
Rmerge	0.134	0.052	0.595
Number of reflections	53950
<I/σ(I)>	5.1	9.5	1.2
Completeness [%]	100.0	99.2	100
Redundancy	3.5	3	3.5

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, HANGING DROP	7.5	293	85 mM HEPES, 8.5% iso-propanol, 17% PEG 4000, 15% glycerol, pH 7.5, VAPOR DIFFUSION, HANGING DROP, temperature 293K