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
SASDE23 – Labeled nuclear pore complex protein Nup153 (NUL-Alexa488/Alexa594) without denaturant
Synchrotron SAXS
data from solutions of
Labeled nuclear pore complex protein Nup153 (NUL-Alexa488/Alexa594) 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 5 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).