5UUI

Crystal Structure of Spin-Labeled T77C TNFa

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	APS BEAMLINE 21-ID-F
Synchrotron site	APS
Beamline	21-ID-F
Temperature [K]	100
Detector technology	CCD
Collection date	2016-03-02
Detector	MARMOSAIC 225 mm CCD
Wavelength(s)	0.97872
Spacegroup name	H 3
Unit cell lengths	65.660, 65.660, 84.090
Unit cell angles	90.00, 90.00, 120.00

Refinement procedure

Resolution	32.830 - 1.400
R-factor	0.1478
Rwork	0.139
R-free	0.17000
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	1tnf
RMSD bond length	0.011
RMSD bond angle	1.274
Data reduction software	XDS
Data scaling software	XSCALE
Phasing software	PHASER
Refinement software	PHENIX ((dev_2356: ???))

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	32.830	1.430
High resolution limit [Å]	1.400	1.400
Number of reflections	26091	1816
<I/σ(I)>	12.71
Completeness [%]	98.0	91.8
Redundancy	3.78	3.02
CC(1/2)	0.997	0.608

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, SITTING DROP	8.5	289	TNFa (VCID10616, B114005) at 10 mg/ml (in 10 mM HEPES, pH = 7.5, 150 mM NaCl) was mixed with an equal volume of protein solution and a solution containing 24% (w/v) PEG-4000, 0.24 M MgCl2, 0.05% DDM, and 0.1 M HEPES/NaOH, pH=8.5. Crystals were produced by sitting drop vapor diffusion at 16 degrees Celsius. Crystals were then soaked with the same solution supplemented with 10% MTSL for one week and harvested with paraffin oil.