Mechanism of Epac activation: structural and functional analyses of Epac2 hinge mutants with constitutive and reduced activities.

Tsalkova T, Blumenthal DK, Mei FC, White MA, Cheng X, J Biol Chem 284(35):23644-51 (2009) Europe PMC

SASDH72 – Isoform 3 of Rap guanine nucleotide exchange factor 4, aE2G-DB08

Isoform 3 of Rap guanine nucleotide exchange factor 4
MWexperimental 114 kDa
MWexpected 114 kDa
VPorod 151 nm3
log I(s) 2.51×102 2.51×101 2.51×100 2.51×10-1
Isoform 3 of Rap guanine nucleotide exchange factor 4 small angle scattering data  s, nm-1
ln I(s)
Isoform 3 of Rap guanine nucleotide exchange factor 4 Guinier plot ln 2.51×102 Rg: 3.8 nm 0 (3.8 nm)-2 s2
(sRg)2I(s)/I(0)
Isoform 3 of Rap guanine nucleotide exchange factor 4 Kratky plot 1.104 0 √3 sRg
p(r)
Isoform 3 of Rap guanine nucleotide exchange factor 4 pair distance distribution function Rg: 3.8 nm 0 Dmax: 12.5 nm

Data validation

Fits and models

log I(s)
 s, nm-1
Isoform 3 of Rap guanine nucleotide exchange factor 4, aE2G-DB08 Rg histogram Rg, nm
SAXS data from solutions of Rap guanine nucleotide exchange factor 4 (aE2G-DB08) in 150 mM NaCl, 1 mM EDTA, 1 mM DTT, and 10 mM Tris-HCl, pH 7.5 were collected using an Anton Paar SAXSess instrument at the Department of Chemistry (University of Utah, USA) with 2D position-sensitive image plates at a sample-detector distance of 0.3 m and at a wavelength of λ = 0.1542 nm (I(s) vs s, where s = 4πsinθ/λ, and 2θ is the scattering angle). One solute concentration of 4.00 mg/ml was measured at 12°C. One 6 hour frame was collected. The data were normalized to the intensity of the transmitted beam and linearly integrated; the scattering of the solvent-blank was subtracted.

An N-terminal expression tag (GSPGIPG) in included in the protein construct.

Isoform 3 of Rap guanine nucleotide exchange factor 4 (Epac2)
Mol. type   Protein
Organism   Mus musculus
Olig. state   Monomer
Mon. MW   114.1 kDa
 
UniProt   Q9EQZ6-3 (1-993)
Sequence   FASTA