Decoupling of size and shape fluctuations in heteropolymeric sequences reconciles discrepancies in SAXS vs. FRET measurements.

Fuertes G, Banterle N, Ruff KM, Chowdhury A, Mercadante D, Koehler C, Kachala M, Estrada Girona G, Milles S, Mishra A, Onck PR, Gräter F, Esteban-Martín S, Pappu RV, Svergun DI, Lemke EA, Proc Natl Acad Sci U S A 114(31):E6342-E6351 (2017) Europe PMC

SASDEB3 – Unlabeled nuclear pore complex protein Nup98-Nup96 (N98) without denaturant

Nuclear pore complex protein Nup98-Nup96
MWexperimental 20 kDa
MWexpected 15 kDa
VPorod 27 nm3
log I(s) 6.12×103 6.12×102 6.12×101 6.12×100
Nuclear pore complex protein Nup98-Nup96 small angle scattering data  s, nm-1
ln I(s)
Nuclear pore complex protein Nup98-Nup96 Guinier plot ln 6.12×103 Rg: 2.9 nm 0 (2.9 nm)-2 s2
(sRg)2I(s)/I(0)
Nuclear pore complex protein Nup98-Nup96 Kratky plot 1.104 0 3 sRg
p(r)
Nuclear pore complex protein Nup98-Nup96 pair distance distribution function Rg: 3.1 nm 0 Dmax: 10.4 nm

Data validation


There are no models related to this curve.

Synchrotron SAXS data from solutions of Unlabeled nuclear pore complex protein Nup98-Nup96 (N98) without denaturant in PBS, 10 mM DTT, pH 7.4 were collected on the EMBL P12 beam line at the PETRA III storage ring (DESY; Hamburg, Germany) using a Pilatus 2M 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 1 and 10 mg/ml were measured at 23°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. The low angle data collected at lower concentrations were extrapolated to infinite dilution and merged with the higher concentration data to yield the final composite scattering curve.

The protein contains a penultimate non-canonical amino acid p-acetylphenylalanine (207 Da) that is represented as U (selenocysteine, 168 Da) in the amino acid sequence for the entry. Therefore, the calculated MW from sequence (MW(expected)) must be adjusted accordingly (ca. 40 Da).

Tags: idp
Nuclear pore complex protein Nup98-Nup96 (N98)
Mol. type   Protein
Organism   Homo sapiens
Olig. state   Monomer
Mon. MW   14.8 kDa
 
UniProt   P52948 (2-150)
Sequence   FASTA