Optical and Structural Characterization of a Chronic Myeloid Leukemia DNA Biosensor.

Cordeiro M, Otrelo-Cardoso AR, Svergun DI, Konarev PV, Lima JC, Santos-Silva T, Baptista PV, ACS Chem Biol 13(5):1235-1242 (2018) Europe PMC

SASDCL5 – Light encoded DNA biosensor: e14Ae13Be14C DNA

e14Ae13Be14C
MWexperimental 35 kDa
MWexpected 31 kDa
VPorod 42 nm3
log I(s) 7.32×101 7.32×100 7.32×10-1 7.32×10-2
e14Ae13Be14C small angle scattering data  s, nm-1
ln I(s)
e14Ae13Be14C Guinier plot ln 7.33×101 Rg: 3.4 nm 0 (3.4 nm)-2 s2
(sRg)2I(s)/I(0)
e14Ae13Be14C Kratky plot 1.104 0 √3 sRg
p(r)
e14Ae13Be14C pair distance distribution function Rg: 3.8 nm 0 Dmax: 14 nm

Data validation

Fits and models

log I(s)
 s, nm-1
e14Ae13Be14C DAMFILT model
Synchrotron SAXS data from solutions of Light encoded DNA biosensor: e14Ae13Be14C DNA in 154 mM NaCl, pH 8.3 were collected at the BM29 beam line at the ESRF (Grenoble, France) using a Pilatus 1M detector at a sample-detector distance of 3 m and at a wavelength of λ = 0.1 nm (I(s) vs s, where s = 4πsinθ/λ and 2θ is the scattering angle). Solute concentrations ranging between 0.3 and 2 mg/ml were measured at 10°C. 20 successive 0.050 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 and the different curves were scaled for protein concentration. The low angle data collected at lower concentration were merged with the highest concentration high angle data to yield the final composite scattering curve. The model depicts the averaged spatial representation of the DNA (DAMFILT occupancy and volume-corrected bead model).

e14Ae13Be14C
Mol. type   DNA
Olig. state   Monomer
Mon. MW   31.4 kDa
Sequence   FASTA