6PJX

Crystal Structure of G Protein-Coupled Receptor Kinase 5 (GRK5) in Complex with Calmodulin (CaM)

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	ROTATING ANODE
Source details	RIGAKU MICROMAX-007 HF
Temperature [K]	93.15
Detector technology	PIXEL
Collection date	2015-12-15
Detector	DECTRIS PILATUS 200K
Wavelength(s)	1.54
Spacegroup name	P 21 21 21
Unit cell lengths	69.875, 83.061, 137.521
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	49.010 - 1.960
R-factor	0.197
Rwork	0.196
R-free	0.23000
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	4tnb
RMSD bond length	0.008
RMSD bond angle	0.901
Data reduction software	HKL-3000
Data scaling software	HKL-3000
Phasing software	PHASER
Refinement software	PHENIX (1.15.2_3472)

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	49.010	2.030
High resolution limit [Å]	1.958	1.960
Rmerge	0.108	0.596
Number of reflections	51717
<I/σ(I)>	14.03	2
Completeness [%]	88.5	63.7
Redundancy	3.9	2.8

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, SITTING DROP	7.5	277.15	Sample- 25 mg/ml GRK5/CaM complex in buffer 20 mM Hepes, pH 7.2, 200 mM NaCl, 2 mM CaCl2, 1 mM DTT incubated with SGV in the presence of MgCl2. Crystallization screening reservoir condition- 90% of (200 mM KCl, 50 mM Hepes pH 7.5, 30% 5/4 PO/OH) + 10% of (0.05% w/v L-Citrulline, 0.05% w/v Glycine, 0.05% w/v L-(-)-Threonine, 0.05% w/v L-(+)-Lysine, 0.05% w/v L-Alanine, 0.05% w/v L-Arginine, 0.05% w/v L-Asparagine monohydrate, 0.05% w/v L-Aspartic acid, 0.05% w/v L-Glutamic acid, 0.05% w/v L-Glutamine, 0.05% w/v L-Histidine, 0.05% w/v L-Isoleucine, 0.05% w/v L-Leucine, 0.05% w/v L-Methionine, 0.05% w/v L-Phenylalanine, 0.05% w/v L-Proline, 0.05% w/v L-Serine, 0.05% w/v L-Tryptophan, 0.05% w/v L-Tyrosine, 0.05% w/v L-Valine, 0.02 M HEPES sodium pH 6)