1GPW

Structural evidence for ammonia tunneling across the (beta/alpha)8 barrel of the imidazole glycerol phosphate synthase bienzyme complex.

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	EMBL/DESY, HAMBURG BEAMLINE X11
Synchrotron site	EMBL/DESY, HAMBURG
Beamline	X11
Temperature [K]	100
Detector technology	CCD
Detector	X-RAY RESEARCH
Spacegroup name	P 1 21 1
Unit cell lengths	74.000, 91.900, 131.000
Unit cell angles	90.00, 105.90, 90.00

Refinement procedure

Resolution	30.000 - 2.400
R-factor	0.226
Rwork	0.226
R-free	0.29000
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	1thf
RMSD bond length	0.008
RMSD bond angle	1.500
Data reduction software	DENZO
Data scaling software	SCALEPACK
Phasing software	AMoRE
Refinement software	CNS (1.0)

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	30.000	2.450
High resolution limit [Å]	2.370	2.370
Rmerge	0.048	0.168
Number of reflections	65356	5485 *
<I/σ(I)>	28	4.5
Completeness [%]	96.7	78.8
Redundancy	2.9	2.5

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	Vapor diffusion *	8		PROTEIN SOLUTION: 10 MM TRIS (PH 8.0), 1 MM DTT, 1 MM EDTA, 28.8 MG/ML PROTEIN COMPLEX. PRECIPITATE SOLUTION: 15 %[W/V] PEG-8000, 0.9 M AMMONIUM NITRATE, 0.1 M HEPES/HCL (PH 8.5), 10 MM DTT, 5% [V/V] MPD

Crystallization Reagents in Literatures

ID	crystal ID	solution	reagent name	concentration (unit)	details
1	1	drop	Tris-NaOH	10 (mM)	pH8.0
2	1	drop		150 (mM)
3	1	drop	EDTA	1 (mM)
4	1	drop	dithiothreitol	5 (mM)
5	1	drop	protein	12.5 (mg/ml)
6	1	reservoir	sodium acetate	0.1 (M)	pH5.0
7	1	reservoir	dithiothreitol	10 (mM)
8	1	reservoir		15 (mM)
9	1	reservoir	PEG200	30 (%(v/v))