A genetically encoded anomalous SAXS ruler to probe the dimensions of intrinsically disordered proteins.

Yu M, Gruzinov AY, Ruan H, Scheidt T, Chowdhury A, Giofrè S, Mohammed ASA, Caria J, Sauter PF, Svergun DI, Lemke EA, Proc Natl Acad Sci U S A 121(50):e2415220121 (2024) Europe PMC

SASDUL9 – Wild-type Importin Beta Binding domain (IBB) from importin subunit alpha-1 in 0.5 M Urea

Importin subunit alpha-1
MWexperimental 14 kDa
MWexpected 9 kDa
VPorod 23 nm3
log I(s) 9.75×10-3 9.75×10-4 9.75×10-5 9.75×10-6
Importin subunit alpha-1 small angle scattering data  s, nm-1
ln I(s)
Importin subunit alpha-1 Guinier plot ln 9.76×10-3 Rg: 2.6 nm 0 (2.6 nm)-2 s2
(sRg)2I(s)/I(0)
Importin subunit alpha-1 Kratky plot 1.104 0 3 sRg
p(r)
Importin subunit alpha-1 pair distance distribution function Rg: 2.8 nm 0 Dmax: 11.6 nm

Data validation


There are no models related to this curve.

Synchrotron SAXS data from solutions of wild-type IBB in phosphate buffered saline containing 0.5 M urea, 0.3 M KCl, 5 mM DTT, pH 7.4 were collected on the EMBL P12 beam line at PETRA III (DESY; Hamburg, Germany) using a Pilatus 6M detector at a sample-detector distance of 3.1 m and at a wavelength of λ = 0.092 nm (I(s) vs s, where s = 4πsinθ/λ, and 2θ is the scattering angle). In-line size-exclusion chromatography (SEC) SAS was employed. The SEC parameters were as follows: A 40.00 μl sample at 20 mg/ml was injected at a 0.40 ml/min flow rate onto a GE Superdex 75 Increase 5/150 column at 20°C. 1800 successive 0.250 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.

Wild-type IBB in 0.5 M urea measured at 13428 eV. SAXS data measured at several different energies spanning the K-absorbtion edge of bromine are made available in the full entry zip archive.

Importin subunit alpha-1 (Wild-type IBB)
Mol. type   Protein
Organism   Homo sapiens
Olig. state   Monomer
Mon. MW   8.7 kDa
 
UniProt   J3QLL0 (2-73)
Sequence   FASTA