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
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).