3I80

Protein Tyrosine Phosphatase 1B - Transition state analog for the second catalytic step

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	ROTATING ANODE
Source details	RIGAKU RU200
Temperature [K]	100
Detector technology	IMAGE PLATE
Collection date	2008-09-06
Detector	RIGAKU RAXIS IV
Wavelength(s)	1.5418
Spacegroup name	P 31 2 1
Unit cell lengths	87.840, 87.840, 103.790
Unit cell angles	90.00, 90.00, 120.00

Refinement procedure

Resolution	35.713 - 2.250
R-factor	0.201
Rwork	0.199
R-free	0.23500
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	1sug
RMSD bond length	0.006
RMSD bond angle	0.939
Data reduction software	d*TREK
Data scaling software	d*TREK (9.4SSI)
Phasing software	PHASER (1.3.3)
Refinement software	PHENIX

Data quality characteristics

	Overall	Inner shell	Outer shell
Low resolution limit [Å]	38.040	38.040	2.330
High resolution limit [Å]	2.250	4.840	2.250
Rmerge	0.084	0.031	0.473
Total number of observations		9378	8938
Number of reflections	22453
<I/σ(I)>	8.6	24.5	2.2
Completeness [%]	100.0	99.8	99.9
Redundancy	4.01	3.9	4.04

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, SITTING DROP	7.5	277	Drop: 2 uL of protein solution, 0.5 uL sucrose 30% (w/v) and 3 uL of precipitant solution (0.1 M HEPES pH 7.5, 0.2 M magnesium acetate and 18-20% polyethylene glycol 8000). Well: 500 uL of precipitant solution. The protein solution was prepared as follows: 9 uL of 50 mM of Na3VO4 and 1 uL of 50 mM of DADEYL peptide (at pH 8.5-9.0) were mixed and allowed to react for 1-1.5 hour; then, 50 uL of native PTP1B (12 mg/mL in 10 mM Tris pH 7.5, 25 mM NaCl, 0.2 mM EDTA and 3 mM DTT) was added and the solution used immediately for crystallization. VAPOR DIFFUSION, SITTING DROP, temperature 277K