4HPU

Crystal structure of the catalytic subunit of cAMP-dependent protein kinase displaying partial phosphoryl transfer of AMP-PNP onto a substrate peptide

Experimental procedure

Source type	SYNCHROTRON
Source details	ALS BEAMLINE 8.2.1
Synchrotron site	ALS
Beamline	8.2.1
Temperature [K]	100
Detector technology	CCD
Detector	ADSC QUANTUM 315r
Wavelength(s)	1.0
Spacegroup name	P 21 21 21
Unit cell lengths	57.690, 79.800, 98.140
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	61.910 - 1.550
R-factor	0.17292
Rwork	0.172
R-free	0.19448
Structure solution method	MOLECULAR REPLACEMENT
RMSD bond length	0.009
RMSD bond angle	1.329
Data reduction software	MOSFLM
Data scaling software	SCALA
Refinement software	REFMAC (5.5.0110)

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	61.910	1.630
High resolution limit [Å]	1.550	1.550
Rmerge	0.076	0.446
Number of reflections	66294
<I/σ(I)>	12.8	3.2
Completeness [%]	99.7	99.2
Redundancy	5.7	5.7

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, HANGING DROP	8	277.15	50 mM Bicine, 150 mM Ammonium acetate, 10 mM DTT, ~7-10 mg/mL protein Well solution: 2% MPD in a 1 mL well. 80 uL of methanol was added to the well immediately before sealing. Drop was 8 uL of 1:1 protein:well, VAPOR DIFFUSION, HANGING DROP, temperature 277.15K, pH 8.0