Allosteric activation of the nitric oxide receptor soluble guanylate cyclase mapped by cryo-electron microscopy.

Horst BG, Yokom AL, Rosenberg DJ, Morris KL, Hammel M, Hurley JH, Marletta MA, Elife 8 (2019) Europe PMC

SASDHA2 – Soluble guanylate cyclase (unbound)

Soluble guanylyl cyclase alpha-1 subunit
Soluble guanylyl cyclase beta-1 subunit
MWexperimental 140 kDa
MWexpected 147 kDa
VPorod 230 nm3
log I(s) 4.19×102 4.19×101 4.19×100 4.19×10-1
Soluble guanylyl cyclase alpha-1 subunit Soluble guanylyl cyclase beta-1 subunit small angle scattering data  s, nm-1
ln I(s)
Soluble guanylyl cyclase alpha-1 subunit Soluble guanylyl cyclase beta-1 subunit Guinier plot ln 4.19×102 Rg: 4.3 nm 0 (4.3 nm)-2 s2
(sRg)2I(s)/I(0)
Soluble guanylyl cyclase alpha-1 subunit Soluble guanylyl cyclase beta-1 subunit Kratky plot 1.104 0 3 sRg
p(r)
Soluble guanylyl cyclase alpha-1 subunit Soluble guanylyl cyclase beta-1 subunit pair distance distribution function Rg: 4.4 nm 0 Dmax: 13.3 nm

Data validation


Fits and models


log I(s)
 s, nm-1
Soluble guanylyl cyclase alpha-1 subunit Soluble guanylyl cyclase beta-1 subunit BILBOMD model

Synchrotron SAXS data from solutions of guanylate cyclase in 50 mM KH2PO4, 150 mM NaCl, 2% glycerol, pH 7.4 were collected on the 12.3.1 (SIBYLS) beam line at the Advanced Light Source (ALS; Berkeley, CA, USA) using a Pilatus3 X 2M detector at a sample-detector distance of 2.1 m and at a wavelength of λ = 0.1127 nm (I(s) vs s, where s = 4πsinθ/λ, and 2θ is the scattering angle). In-line size-exclusion chromatography (SEC) SAS was employed. The SEC parameters were as follows: A 60.00 μl sample at 7 mg/ml was injected at a 0.45 ml/min flow rate onto a Shodex KW-800 series column at 20°C. 20 successive 3 second frames were collected through the SEC elution peak of the sample. The data were normalized to the intensity of the transmitted beam and radially averaged; the scattering of the solvent-blank was subtracted.

Soluble guanylyl cyclase alpha-1 subunit
Mol. type   Protein
Organism   Manduca sexta
Olig. state   Monomer
Mon. MW   78.5 kDa
 
UniProt   O77105 (1-699)
Sequence   FASTA
 
Soluble guanylyl cyclase beta-1 subunit
Mol. type   Protein
Organism   Manduca sexta
Olig. state   Monomer
Mon. MW   68.1 kDa
 
UniProt   O77106 (1-600)
Sequence   FASTA