4KYT

The structure of superinhibitory phospholamban bound to the calcium pump SERCA1a

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	APS BEAMLINE 19-ID
Synchrotron site	APS
Beamline	19-ID
Temperature [K]	100
Detector technology	CCD
Collection date	2013-02-27
Detector	ADSC QUANTUM 315
Wavelength(s)	0.98
Spacegroup name	P 21 21 21
Unit cell lengths	61.591, 93.094, 316.375
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	50.000 - 2.833
R-factor	0.24139
Rwork	0.239
R-free	0.28388
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	2c8l
RMSD bond length	0.009
RMSD bond angle	1.196
Data reduction software	HKL-3000
Data scaling software	HKL-3000
Phasing software	PHASER
Refinement software	REFMAC (5.5.0109)

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	50.000	2.890
High resolution limit [Å]	2.833	2.833
Rmerge	0.084	0.492
Number of reflections	42992
<I/σ(I)>	14.4	2.6
Completeness [%]	97.2	97.3
Redundancy	5.5	4.5

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, HANGING DROP		277	1 ul of SERCA1a at 15 mg/mL in 2% n-nonyl-beta-D-maltopyranoside (nonyl maltoside) (Anatrace), 20% glycerol, 100 mM MOPS (pH 7.0), 0.12 M sucrose, 80 mM KCl, 3 mM MgCl2, and 2.8 mM EGTA was mixed with 1 uL of phospholamban at 2.1 mg/mL in 20 mM MOPS (pH 7.2), 20% glycerol, and 0.1 % decyl maltoside or 0.01% dodecyl maltoside. This protein mixture was then added to an equal volume of crystallization liquor; 15 % glycerol, 17% (W/V) PEG-2000, 200mM NaOAc, and 5 mM beta-mercoptoethanol), VAPOR DIFFUSION, HANGING DROP, temperature 277K