3TCM

Crystal Structure of Alanine Aminotransferase from Hordeum vulgare

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	APS BEAMLINE 22-ID
Synchrotron site	APS
Beamline	22-ID
Temperature [K]	100
Detector technology	CCD
Collection date	2009-02-26
Detector	MARMOSAIC 300 mm CCD
Wavelength(s)	1.0
Spacegroup name	P 21 21 2
Unit cell lengths	119.859, 126.973, 75.663
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	87.160 - 2.710
R-factor	0.21779
Rwork	0.215
R-free	0.27904
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	The phasing template was prepared by the CCP4 program chainsaw from the Pyrococcus furiosis Pfu-1397077-001 (PDB entry 1xi9) coordinate file and the Hv (i.e. barley) AlaAT-PfAlaAT sequence alignment.
RMSD bond length	0.029
RMSD bond angle	2.449
Data reduction software	HKL-2000
Data scaling software	HKL-2000
Phasing software	PHASER (& RESOLVE)
Refinement software	REFMAC (5.5.0102)

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	87.200	2.800
High resolution limit [Å]	2.710	2.700
Rmerge	0.100	0.493
Number of reflections	31088
<I/σ(I)>	15	2.46
Completeness [%]	97.1	98.8
Redundancy	4.1	4.1

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, SITTING DROP	6	295	The protein sample was 12 mg/mL in 20 mM carbonate buffer-pH 10.5, 2.5 mM PLP, 2.5 mM cycloserine, and 2.5 mM DTT. Crystals were obtained using a 0.7 mL well composition of 22(w/v)% polyethylene glycol monomethyl ether (PGME) 5000, 0.1 M MES, 9 (v/v) % tacsimate and a 2 uL sitting drop prepared from equal volumes of protein and well solution., VAPOR DIFFUSION, SITTING DROP, temperature 295K