Open and extended conformation of Human DNA ligase-1

farheen jahan.

SASDV36 – Human DNA ligase-1 (apo form)

DNA ligase 1
MWexperimental 100 kDa
MWexpected 102 kDa
VPorod 53 nm3
log I(s) 1.29×104 1.29×103 1.29×102 1.29×101
DNA ligase 1 small angle scattering data  s, nm-1
ln I(s)
DNA ligase 1 Guinier plot ln 1.30×104 Rg: 3.6 nm 0 (3.6 nm)-2 s2
(sRg)2I(s)/I(0)
DNA ligase 1 Kratky plot 1.104 0 √3 sRg
p(r)
DNA ligase 1 pair distance distribution function Rg: 4.2 nm 0 Dmax: 18.4 nm

Data validation

Fits and models

log I(s)
 s, nm-1
DNA ligase 1 DAMMIF model
SAXS data from solutions of DNA ligase-1 in 50 mM HEPES pH 7.5, 200 mM NaCl were collected using an Anton Pair SAXSpace instrument at the CSIR-Central Drug Research Institute (Lucknow, India) equipped with a Mythen2 R 1K detector at a sample-detector distance of 0.3 m and at a wavelength of λ = 0.154 nm (I(s) vs s, where s = 4πsinθ/λ, and 2θ is the scattering angle). One solute concentration of 2.00 mg/ml was measured at 10°C. Two successive 30 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.

CAUTION!! The Porod volume (~53 nm³) and molecular weight estimates calculated from the SAXS data (22-43 kDa) in addition to the volume (~56 nm³) and molecular weight of the DAM model (~28 kDa) are significantly lower than expected for a protein of 100 kDa.

DNA ligase 1
Mol. type   Protein
Organism   Homo sapiens
Olig. state   Monomer
Mon. MW   101.7 kDa
Sequence   FASTA