Conformational analysis of a genetically encoded FRET biosensor by SAXS.

Mertens HD, Piljić A, Schultz C, Svergun DI, Biophys J 102(12):2866-75 (2012) Europe PMC

SASDAF5 – CYNEX4-T266D

CYNEX4 FRET probe, (eYFP-AnnexinA4-eCFP) T266D mutant
MWI(0) 96 kDa
MWexpected 93 kDa
VPorod 146 nm3
log I(s) 8.80×101 8.80×100 8.80×10-1 8.80×10-2
CYNEX4 FRET probe, (eYFP-AnnexinA4-eCFP) T266D mutant small angle scattering data  s, nm-1
ln I(s)
CYNEX4 FRET probe, (eYFP-AnnexinA4-eCFP) T266D mutant Guinier plot ln 8.80×101 Rg: 4.1 nm 0 (4.1 nm)-2 s2
(sRg)2I(s)/I(0)
CYNEX4 FRET probe, (eYFP-AnnexinA4-eCFP) T266D mutant Kratky plot 1.104 0 √3 sRg
p(r)
CYNEX4 FRET probe, (eYFP-AnnexinA4-eCFP) T266D mutant pair distance distribution function Rg: 4.2 nm 0 Dmax: 14.4 nm

Data validation

Fits and models

log I(s)
 s, nm-1
CYNEX4 FRET probe, (eYFP-AnnexinA4-eCFP) T266D mutant DAMMIF model
log I(s)
 s, nm-1
CYNEX4 FRET probe, (eYFP-AnnexinA4-eCFP) T266D mutant CORAL model
Synchrotron SAXS data from solutions of CYNEX4-T266D in 50 mM HEPES 50 mM KCl, pH 7.5 were collected on the EMBL X33 beam line at the DORIS III, DESY storage ring (Hamburg, Germany) using a Pilatus 1M-W detector at a sample-detector distance of 2.7 m and at a wavelength of λ = 0.15 nm (I(s) vs s, where s = 4πsinθ/λ, and 2θ is the scattering angle). Solute concentrations ranging between 0.9 and 17 mg/ml were measured at 10°C. Eight successive 15 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 low angle data collected at lower concentration were merged with the highest concentration high angle data to yield the final composite scattering curve.

PED: https://proteinensemble.org/PED00010

Tags: X33 PED
CYNEX4 FRET probe, (eYFP-AnnexinA4-eCFP) T266D mutant (CYNEX4-T266D)
Mol. type   Protein
Organism   Homo sapiens
Olig. state   Monomer
Mon. MW   92.8 kDa
Sequence   FASTA
 
PED ID   PED00010