1Q4A

S65T Q80R Green Fluorescent Protein (GFP) pH 8.5

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	ROTATING ANODE
Source details	RIGAKU
Detector technology	IMAGE PLATE
Detector	RIGAKU RAXIS IV
Spacegroup name	P 21 21 21
Unit cell lengths	50.864, 62.351, 69.174
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	25.000 - 1.450
R-factor	0.187
Rwork	0.187
R-free	0.20100
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	1ema
RMSD bond length	0.010 *
RMSD bond angle	1.740 *
Data reduction software	DENZO
Data scaling software	SCALEPACK
Phasing software	CNS
Refinement software	CNS

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	25.000
High resolution limit [Å]	1.450
Rmerge	0.041 *	0.302 *
Number of reflections	36969
Completeness [%]	93.3	68.5 *
Redundancy	2.8 *

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, HANGING DROP	8.5	298	PEG 4000, MgCl2, BME, pH 8.5, VAPOR DIFFUSION, HANGING DROP, temperature 298K

Crystallization Reagents in Literatures

ID	crystal ID	solution	reagent name	concentration (unit)	details
1	1	reservoir	PEG4000	22-26 (%)
2	1	reservoir		50 (mM)
3	1	reservoir	2-mercaptoethanol	10 (mM)
4	1	reservoir	HEPES	50 (mM)	pH8.1-8.5
5	1	drop	protein	12-15 (mg/ml)
6	1	drop	HEPES	50 (mM)	pH7.5