4I6O

Crystal structure of chemically synthesized human anaphylatoxin C3a

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	AUSTRALIAN SYNCHROTRON BEAMLINE MX2
Synchrotron site	Australian Synchrotron
Beamline	MX2
Temperature [K]	100
Detector technology	CCD
Collection date	2012-10-24
Detector	ADSC QUANTUM 315r
Wavelength(s)	0.95370
Spacegroup name	P 63
Unit cell lengths	68.285, 68.285, 39.266
Unit cell angles	90.00, 90.00, 120.00

Refinement procedure

Resolution	19.420 - 2.140
R-factor	0.21839
Rwork	0.208
R-free	0.26793
Structure solution method	MOLECULAR REPLACEMENT
RMSD bond length	0.017
RMSD bond angle	1.907
Data reduction software	MOSFLM
Data scaling software	SCALA
Phasing software	PHASER
Refinement software	REFMAC (5.7.0029)

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	39.270	2.200
High resolution limit [Å]	2.140	2.140
Rmerge	0.106	0.787
Number of reflections	5870
<I/σ(I)>	15.9	3.4
Completeness [%]	100.0	100
Redundancy	22.1	22

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, HANGING DROP	8.5	277	VAPOR DIFFUSION, HANGING DROP, 1 uL 5 mg/mL lyophilized C3a in 2 mM hydrochloric acid + 1 uL reservoir (0.1 M Tris-HCl, 0.2 M ammonium phosphate monobasic, 49-51% v/v MPD, pH 8.5) against 500 uL reservoir, prepared at 277K, incubated at 281K for 96 hours, stored at 277K, rhomboid-shaped crystals appeared after 6 days and grew to a final size of 0.5 x 0.2 x 0.2 mm