4JC6

Mercury activation of the plant aquaporin SoPIP2;1 - structural and functional characterization

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	NSLS BEAMLINE X25
Synchrotron site	NSLS
Beamline	X25
Temperature [K]	100
Detector technology	CCD
Collection date	2008-04-10
Detector	ADSC QUANTUM 315
Wavelength(s)	0.9795
Spacegroup name	P 21 21 21
Unit cell lengths	123.546, 141.818, 186.515
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	29.900 - 2.152
R-factor	0.19843
Rwork	0.197
R-free	0.22704
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	1z98
RMSD bond length	0.012
RMSD bond angle	1.333
Data reduction software	DENZO
Data scaling software	SCALEPACK
Phasing software	MOLREP
Refinement software	REFMAC

Data quality characteristics

	Overall	Inner shell	Outer shell
Low resolution limit [Å]	30.000	30.000	2.230
High resolution limit [Å]	2.150	4.630	2.150
Rmerge	0.129	0.069	0.639
Number of reflections	176544
<I/σ(I)>	5.9
Completeness [%]	99.8	99.9	98.9
Redundancy	6	5.9	5.6

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, HANGING DROP	7	280	The protein was incubated with 5mM HgCl2, and excess HgCl2 was washed away. Crystallisation condition was 26% PEG400, 25mM Tris pH 7.0, 100mM NaCl. The precipitant solution was mixed with 1/5 0.1M CdCl2. Protein and precipitant solution was mixed in 1:1 ratio , VAPOR DIFFUSION, HANGING DROP, temperature 280K