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

SASDEH2 – Unlabeled nucleoporin NUP49/NSP49 (N49) without denaturant

Nucleoporin NUP49/NSP49
MWexperimental 4 kDa
MWexpected 4 kDa
VPorod 4 nm3
log I(s) 7.61×101 7.61×100 7.61×10-1 7.61×10-2
Nucleoporin NUP49/NSP49 small angle scattering data  s, nm-1
ln I(s)
Nucleoporin NUP49/NSP49 Guinier plot ln 7.61×101 Rg: 1.6 nm 0 (1.6 nm)-2 s2
(sRg)2I(s)/I(0)
Nucleoporin NUP49/NSP49 Kratky plot 1.104 0 3 sRg
p(r)
Nucleoporin NUP49/NSP49 pair distance distribution function Rg: 1.7 nm 0 Dmax: 6 nm

Data validation


Fits and models


log I(s)
 s, nm-1
Unlabeled nucleoporin NUP49/NSP49 (N49) without denaturant Rg histogram Rg, nm
Nucleoporin NUP49/NSP49 OTHER model
Nucleoporin NUP49/NSP49 OTHER model
Nucleoporin NUP49/NSP49 OTHER model
Nucleoporin NUP49/NSP49 OTHER model
Nucleoporin NUP49/NSP49 OTHER model
Nucleoporin NUP49/NSP49 OTHER model
Nucleoporin NUP49/NSP49 OTHER model
Nucleoporin NUP49/NSP49 OTHER model
Nucleoporin NUP49/NSP49 OTHER model

Synchrotron SAXS data from solutions of Unlabeled nucleoporin NUP49/NSP49 (N49) 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 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 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 NUP49/NSP49 (N49)
Mol. type   Protein
Organism   Saccharomyces cerevisiae
Olig. state   Monomer
Mon. MW   3.9 kDa
 
UniProt   Q02199 (121-154)
Sequence   FASTA