An N-terminally truncated form of cyclic GMP-dependent protein kinase Iα (PKG Iα) is monomeric and autoinhibited and provides a model for activation.

Moon TM, Sheehe JL, Nukareddy P, Nausch LW, Wohlfahrt J, Matthews DE, Blumenthal DK, Dostmann WR, J Biol Chem 293(21):7916-7929 (2018) PubMed

SASDDS4 – cGMP-dependent protein kinase 1: ∆53 PKG Iα

cGMP-dependent protein kinase 1
MWI(0) 76 kDa
MWexpected 70 kDa
VPorod 105 nm3
log I(s) 3.21×102 3.21×101 3.21×100 3.21×10-1
cGMP-dependent protein kinase 1 small angle scattering data  s, nm-1
ln I(s)
cGMP-dependent protein kinase 1 Guinier plot ln 3.21×102 Rg: 3.0 nm 0 (3.0 nm)-2 s2
(sRg)2I(s)/I(0)
cGMP-dependent protein kinase 1 Kratky plot 1.104 0 3 sRg
p(r)
cGMP-dependent protein kinase 1 pair distance distribution function Rg: 3.0 nm 0 Dmax: 9.7 nm

Experimental data validation


Fits and models


log I(s)
 s, nm-1
cGMP-dependent protein kinase 1 DAMFILT model
Synchrotron SAXS data from solutions of cGMP-dependent protein kinase 1 (∆53 PKG Iα) were collected using SEC-SAXS on the BL4-2 beam line at the Stanford Synchrotron Radiation Lightsource (SSRL; Stanford, CA, USA) using a Rayonix MX225-HE detector at a sample-detector distance of 1.7 m and at a wavelength of λ = 0.1127 nm (l(s) vs s, where s = 4πsinθ/λ, and 2θ is the scattering angle). A 35 μL sample of 6 mg/mL PKG Iα ∆53 was injected onto a Superdex 200 3.2/30 column (GE) and eluted isocratically using an Ettan FPLC (GE) at 0.05 mL/min in 50 mM MES, 300 mM NaCl, 1mM TCEP, pH 6.9 supplemented with 5 mM DTT as a radical scavenger. The eluant stream was connected in-line to a 1.5 mm quartz capillary. Data were collected with a 1 second/frame exposure rate at 22°C. A total of 600 frames were collected. Frames 410-459 containing the peak corresponding to ∆53 PKG were averaged and subtracted from background. The scattering curve, Guinier and P(r) functions, and Porod volume were calculated using the PRIMUS (ATSAS) program suite.

cGMP-dependent protein kinase 1 (∆53 PKG Iα)
Mol. type   Protein
Organism   Bos taurus
Olig. state   Monomer
Mon. MW   70.4 kDa
 
UniProt   P00516 (54-671)
Sequence   FASTA