4X6R

An Isoform-specific Myristylation Switch Targets RIIb PKA Holoenzymes to Membranes

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	ALS BEAMLINE 8.2.1
Synchrotron site	ALS
Beamline	8.2.1
Temperature [K]	200
Detector technology	CCD
Collection date	2013-07-04
Detector	ADSC QUANTUM 1
Wavelength(s)	1
Spacegroup name	P 32 2 1
Unit cell lengths	125.760, 125.760, 140.881
Unit cell angles	90.00, 90.00, 120.00

Refinement procedure

Resolution	62.880 - 2.400
R-factor	0.186
Rwork	0.184
R-free	0.23400
RMSD bond length	0.022
RMSD bond angle	1.857
Data reduction software	MOSFLM
Data scaling software	HKL-2000
Phasing software	PHASER
Refinement software	PHENIX ((PHENIX.REFINE: 1.7_650))

Data quality characteristics

	Overall
Low resolution limit [Å]	63.000
High resolution limit [Å]	2.400
Number of reflections	48209
<I/σ(I)>	3
Completeness [%]	99.9
Redundancy	4

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	BATCH MODE		298	Crystallization of RIb(91-379,R333K): myrC(K7C) heterodimer :RC heterodimer that was concentrated to 14 mg/mL and screened against different ammonium sulfate concentrations ranging from 0.8-2.5 M in 0.1 M sodium citrate buffer and also varying the pH from 5.0-6.0 using the hanging drop vapor diffusion method. The crystal used for structure determination was obtained from a 4 uL drop containing 1:1 protein to well solution with the well solution containing 1.6 M ammonium sulfate and 0.1 M sodium citrate at pH 5.5