Membranes prime the RapGEF EPAC1 to transduce cAMP signaling

Sartre C, Peurois F, Ley M, Kryszke M, Zhang W, Courilleau D, Fischmeister R, Ambroise Y, Zeghouf M, Cianferani S, Ferrandez Y, Cherfils J, Nature Communications 14(1) (2023) DOI

SASDQC8 – Rap guanine nucleotide exchange factor Epac1 with CE3F4 inhibitor

Rap guanine nucleotide exchange factor 3
MWexperimental 83 kDa
MWexpected 82 kDa
VPorod 140 nm3
log I(s) 3.05×10-2 3.05×10-3 3.05×10-4 3.05×10-5
Rap guanine nucleotide exchange factor 3 small angle scattering data  s, nm-1
ln I(s)
Rap guanine nucleotide exchange factor 3 Guinier plot ln 3.06×10-2 Rg: 3.3 nm 0 (3.3 nm)-2 s2
(sRg)2I(s)/I(0)
Rap guanine nucleotide exchange factor 3 Kratky plot 1.104 0 √3 sRg
p(r)
Rap guanine nucleotide exchange factor 3 pair distance distribution function Rg: 3.3 nm 0 Dmax: 11.4 nm

Data validation

There are no models related to this curve.

Synchrotron SAXS data from solutions of Rap guanine nucleotide exchange factor Epac1 with CE3F4 inhibitor in 20 mM Tris pH 8.0, 150 mM NaCl, 0.5 mM EDTA were collected on the SWING beam line at SOLEIL (Saint-Aubin, France) using a AVIEX PCCD170170 detector at a wavelength of λ = 0.103 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 50.00 μl sample at 5 mg/ml was injected at a 0.20 ml/min flow rate onto a Agilent Bio SEC-3, 300 Å column at 20°C. 180 successive 0.750 second frames were collected. The data were normalized to the intensity of the transmitted beam and radially averaged; the scattering of the solvent-blank was subtracted.

The CE3F4 inhibitor alone causes virtually no conformational change of Epac1. CE3F4 = 5,7-Dibromo-6-fluoro-3,4-dihydro-2-methyl-1(2H)-quinolinecarboxaldehyde (PubChem ID: 21781066; https://pubchem.ncbi.nlm.nih.gov/compound/21781066)

Rap guanine nucleotide exchange factor 3
Mol. type   Protein
Organism   Homo sapiens
Olig. state   Monomer
Mon. MW   82.1 kDa
 
UniProt   O95398 (194-923)
Sequence   FASTA