9PQC

Room temperature crystal structure of enhanced Green Fluorescent Protein

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	SSRL BEAMLINE BL12-2
Synchrotron site	SSRL
Beamline	BL12-2
Temperature [K]	277
Detector technology	PIXEL
Collection date	2019-12-01
Detector	DECTRIS PILATUS 6M
Wavelength(s)	0.95369
Spacegroup name	P 21 21 21
Unit cell lengths	51.533, 62.830, 70.636
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	46.950 - 1.400
R-factor	0.1208
Rwork	0.119
R-free	0.15630
Structure solution method	MOLECULAR REPLACEMENT
RMSD bond length	0.010
RMSD bond angle	0.988
Data reduction software	DIALS (2.dev.886-gaa85dd3)
Data scaling software	Aimless (0.7.4)
Phasing software	PHENIX (1.14_3260)
Refinement software	PHENIX (1.21.1_5286)

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	46.950	1.430
High resolution limit [Å]	1.400	1.400
Rmerge	0.167
Rmeas	0.173	0.044
Rpim	0.045	1.043
Number of reflections	48817	3008
<I/σ(I)>	9.36	0.94
Completeness [%]	99.8	99.58
Redundancy	14.4	14.7
CC(1/2)	0.999	0.362

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, HANGING DROP	6.5	277	0.5 microliters of 10 mg/mL protein was mixed with 0.5 microliters of 100 mM MES pH 6.5, 300 mM calcium acetate, and 20% (w/v) PEG 8,000 and allowed to equilibrate over a 500 microliter reservoir over several days. Large single crystals from seeding into 100 mM MES pH 6.5, 300 mM calcium acetate, and 4-10% (w/v) PEG 8,000 in the same format.