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 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.

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