4HPT

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

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	ALS BEAMLINE 8.2.2
Synchrotron site	ALS
Beamline	8.2.2
Temperature [K]	100
Wavelength(s)	1.0
Spacegroup name	P 21 21 21
Unit cell lengths	71.545, 79.060, 80.090
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	44.350 - 2.150
R-factor	0.18907
Rwork	0.187
R-free	0.22797
Structure solution method	MOLECULAR REPLACEMENT
RMSD bond length	0.009
RMSD bond angle	1.155
Data reduction software	MOSFLM
Data scaling software	SCALA
Refinement software	REFMAC (5.5.0110)

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	44.350	2.270
High resolution limit [Å]	2.150	2.150
Rmerge	0.100	0.444
Number of reflections	21475
<I/σ(I)>	12.4	2.8
Completeness [%]	85.7	66.2
Redundancy	7.2	3.8

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, HANGING DROP	8	277.15	Protein buffer: 50 mM Bicine, 150 mM Ammonium Acetate, 10 mM DTT, ~7-10 mg/mL protein Well Solution: 1 mL of 2% MPD, 80 uL methanol added to the well immediately before sealing 8 uL drops of 1:1 protein:well were used., VAPOR DIFFUSION, HANGING DROP, temperature 277.15K, pH 8.0