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

SASDEZ2 – Unlabeled nuclear pore complex protein Nup153 (NUL) without denaturant

Nuclear pore complex protein Nup153
MWexperimental 23 kDa
MWexpected 12 kDa
VPorod 32 nm3
log I(s) 6.54×102 6.54×101 6.54×100 6.54×10-1
Nuclear pore complex protein Nup153 small angle scattering data  s, nm-1
ln I(s)
Nuclear pore complex protein Nup153 Guinier plot ln 6.54×102 Rg: 3 nm 0 (3 nm)-2 s2
(sRg)2I(s)/I(0)
Nuclear pore complex protein Nup153 Kratky plot 1.104 0 √3 sRg
p(r)
Nuclear pore complex protein Nup153 pair distance distribution function Rg: 3.2 nm 0 Dmax: 10.9 nm

Data validation

Fits and models

log I(s)
 s, nm-1
Unlabeled nuclear pore complex protein Nup153 (NUL) without denaturant Rg histogram Rg, nm
Nuclear pore complex protein Nup153 OTHER model
Nuclear pore complex protein Nup153 OTHER model
Nuclear pore complex protein Nup153 OTHER model
Nuclear pore complex protein Nup153 OTHER model
Synchrotron SAXS data from solutions of Unlabeled nuclear pore complex protein Nup153 (NUL) 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 2 and 5 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 Nup153 (NUL)
Mol. type   Protein
Organism   Homo sapiens
Olig. state   Monomer
Mon. MW   11.7 kDa
 
UniProt   P49790 (884-993)
Sequence   FASTA