1B9C

Green Fluorescent Protein Mutant F99S, M153T and V163A

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	ELETTRA BEAMLINE 5.2R
Synchrotron site	ELETTRA
Beamline	5.2R
Temperature [K]	100
Detector technology	IMAGE PLATE
Detector	MAR scanner 345 mm plate
Spacegroup name	P 21 21 21
Unit cell lengths	83.480, 85.370, 140.060
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	30.000 - 2.400
R-factor	0.204
Rwork	0.204
R-free	0.28000
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	1gfl
RMSD bond length	0.015
RMSD bond angle	28.480 *
Data reduction software	MOSFLM
Data scaling software	CCP4 ((SCALA))
Phasing software	AMoRE
Refinement software	X-PLOR (3.8.5)

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	30.000	2.640
High resolution limit [Å]	2.370	2.370
Rmerge	0.062	0.146
Total number of observations	165357 *
Number of reflections	36746	5445 *
<I/σ(I)>	6.9	4.2
Completeness [%]	83.9	45
Redundancy	4.6	3.9

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	Vapor diffusion, hanging drop *	7.4 *	20 *	22% PEG 4000, 50 MM HEPES PH 8.5, 50 MM MGCL2, 10 MM 2-MERCAPTOETHANOL, 23% MG/ ML PROTEINA, pH 8.3

Crystallization Reagents in Literatures

ID	crystal ID	solution	reagent name	concentration (unit)	details
1	1	drop	protein	23 (mg/ml)
2	1	drop	HEPES	10 (mM)
3	1	reservoir	PEG4000	22 (%)
4	1	reservoir	HEPES	50 (mM)
5	1	reservoir		50 (mM)
6	1	reservoir	2-mercaptoethanol	10 (mM)