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

SASDEC3 – Unlabeled nucleoporin NSP1 (NSP) without denaturant

Nucleoporin NSP1
MWexperimental 10 kDa
MWexpected 18 kDa
VPorod 45 nm3
log I(s) 8.82×103 8.82×102 8.82×101 8.82×100
Nucleoporin NSP1 small angle scattering data  s, nm-1
ln I(s)
Nucleoporin NSP1 Guinier plot ln 8.83×103 Rg: 4.1 nm 0 (4.1 nm)-2 s2
Nucleoporin NSP1 Kratky plot 1.104 0 3 sRg
Nucleoporin NSP1 pair distance distribution function Rg: 4.1 nm 0 Dmax: 15 nm

Data validation

There are no models related to this curve.

Synchrotron SAXS data from solutions of Unlabeled nucleoporin NSP1 (NSP) without denaturant in PBS, 10 mM DTT, pH 7.4 were collected on the EMBL P12 beam line at the PETRA III storage ring (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 concentration were merged with the highest concentration high angle 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
Nucleoporin NSP1
Mol. type   Protein
Organism   Saccharomyces cerevisiae
Olig. state   Monomer
Mon. MW   17.6 kDa
UniProt   P14907 (2-175)
Sequence   FASTA