4J88

Dark-state structure of sfGFP containing the unnatural amino acid p-azido-phenylalanine at residue 66

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	DIAMOND BEAMLINE I04
Synchrotron site	Diamond
Beamline	I04
Temperature [K]	100
Detector technology	CCD
Collection date	2011-12-04
Detector	ADSC QUANTUM 315r
Wavelength(s)	0.9795
Spacegroup name	P 21 21 21
Unit cell lengths	52.430, 97.560, 102.900
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	48.780 - 2.080
R-factor	0.1684
Rwork	0.165
R-free	0.22820
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	2b3p
RMSD bond length	0.021
RMSD bond angle	1.194
Data reduction software	xia2
Data scaling software	SCALA (3.3.20)
Phasing software	PHASER
Refinement software	REFMAC (5.7.0029)

Data quality characteristics

	Overall	Inner shell	Outer shell
Low resolution limit [Å]	102.900	102.900	2.130
High resolution limit [Å]	2.080	9.300	2.080
Rmerge	0.131	0.035	0.818
Total number of observations		2909	19051
Number of reflections	32505
<I/σ(I)>	13.6	16.4	0.9
Completeness [%]	100.0	99.7	100
Redundancy	7.9	6.6	8.1

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, SITTING DROP	8.3	278	10 mg/mL protein and 100 mM Tris-HCl, pH 8.3, 2.8 M (NH4)2SO4 (200+200 nanoL drop against 60 microL reservoir), VAPOR DIFFUSION, SITTING DROP, temperature 278K