1B48

CRYSTAL STRUCTURE OF MGSTA4-4 IN COMPLEX WITH GSH CONJUGATE OF 4-HYDROXYNONENAL IN ONE SUBUNIT AND GSH IN THE OTHER: EVIDENCE OF SIGNALING ACROSS DIMER INTERFACE IN MGSTA4-4

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	ROTATING ANODE
Source details	ENRAF-NONIUS FR591
Temperature [K]	293
Detector technology	IMAGE PLATE
Collection date	1997-05-15
Detector	MAC Science DIP-2020
Spacegroup name	P 21 21 2
Unit cell lengths	114.450, 96.140, 50.840
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	20.000 - 2.600
R-factor	0.244
Rwork	0.244
R-free	0.31700
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	1guh
RMSD bond length	0.008
RMSD bond angle	19.790 *
Data reduction software	HKL-2000
Data scaling software	HKL-2000
Phasing software	AMoRE
Refinement software	X-PLOR (3.843)

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	20.000	2.690
High resolution limit [Å]	2.600	2.600
Rmerge	0.075	0.256
Number of reflections	15647
<I/σ(I)>	14	2.35
Completeness [%]	87.4	60.1
Redundancy	12.1	2.74

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	Vapor diffusion, hanging drop *	7.5	18-20 *	CRYSTALS WERE GROWN IN HANGING DROPS WHICH INITIALLY CONSISTED OF 0.11 MM PROTEIN IN 0.047 M HEPES BUFFER (PH 7.5) CONTAINING 1.82 MM GLUTATHIONE, 10.91 MM HNA-SG (GSH CONJUGATE OF 4-HYDROXYNONENAL), 9.1% ETHANOL, 4.5% ISOPROPANOL, AND 6.5% PEG MONOMETHYL ETHER 5K (PH 7.5). THE DROPS WERE EQUILIBRATED AT 293 K AGAINST WELL SOLUTION CONTAINING 10% ISOPROPANOL AND 12% PEG MONOMETHYL ETHER 5K IN 80 MM HEPES BUFFER (PH 7.5).

Crystallization Reagents in Literatures

ID	crystal ID	solution	reagent name	concentration (unit)	details
1	1	drop	protein	0.11 (mM)
10	1	reservoir	HEPES	0.08 (M)
11	1	drop	GSHna	11 (mM)
2	1	drop	GSH	1.8 (mM)
3	1	drop	HEPES	3.6 (mM)
4	1	drop	ethanol		1ml
5	1	drop	2-propanol	5.5 (%)
6	1	drop	mPEG5000	6.5 (%)
7	1	drop	HEPES	0.044 (M)
8	1	reservoir	2-propanol	10 (%)
9	1	reservoir	mPEG5000	12 (%)