4WBB

Single Turnover Autophosphorylation Cycle of the PKA RIIb Holoenzyme

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]	200
Detector technology	CCD
Collection date	2012-09-30
Detector	ADSC QUANTUM 1
Wavelength(s)	1
Spacegroup name	C 2 2 2
Unit cell lengths	150.380, 213.951, 61.967
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	50.000 - 2.800
Rwork	0.232
R-free	0.27900
Phasing software	PHASER

Data quality characteristics

	Overall
Low resolution limit [Å]	50.000
High resolution limit [Å]	2.800
Number of reflections	23056
<I/σ(I)>	23.6
Completeness [%]	91.7
Redundancy	3.5

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	BATCH MODE		298	The RIIb (R230K):C apo complex was crystallized in 20%PEG8000, 8% Ethylene Glycol, 20mM MESpH7.5 HEPES . To make CaATP binding holoenzyme crystals, the apo holoenzyme crystals were soaked with 20%PEG8000, 8% Ethylene Glycol, 20mM MESpH7.5 HEPES, 5mM CaCl2 and 1mM ATP at pH5.8 solution overnight