5UM4

Crystal structure of the F255A mutant Kir3.1 cytoplasmic pore domain

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	ALS BEAMLINE 8.2.1
Synchrotron site	ALS
Beamline	8.2.1
Temperature [K]	100
Detector technology	CCD
Collection date	2015-10-26
Detector	ADSC QUANTUM 315r
Wavelength(s)	1.0003
Spacegroup name	P 4 21 2
Unit cell lengths	80.070, 80.070, 85.040
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	85.040 - 2.500
R-factor	0.2263
Rwork	0.224
R-free	0.26670
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	1n9p
RMSD bond length	0.010
RMSD bond angle	1.137
Data reduction software	MOSFLM
Data scaling software	SCALA (3.3.22)
Phasing software	PHASER (2.6.1)
Refinement software	PHENIX

Data quality characteristics

	Overall	Inner shell	Outer shell
Low resolution limit [Å]	85.040	7.910	2.640
High resolution limit [Å]	1.987	5.590	2.500
Rmerge		0.040	0.591
Rmeas	0.067	0.042	0.615
Rpim	0.019	0.012	0.169
Total number of observations	125290
Number of reflections	10030
<I/σ(I)>	21.9	15.4	1.3
Completeness [%]	99.6	100	100
Redundancy	12.5	11.6	13.2

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, HANGING DROP	5	277	The protein was concentrated to 10 mg/mL. Crystals grown by mixing equal amounts of protein and reservoir solution (25 mM Na/K phosphate pH 5.0, 40 mM NaCl, 35 % Peg 400), and allowed to equilibrate over 1 ml of reservoir solution.