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

SASDUU9 – Br-labelled Elastin-like Polypeptide (ELP) in 2 M Urea

Elastin-like polypeptide labeled with unnatural amino acid diBrK
MWexperimental 9 kDa
MWexpected 13 kDa
VPorod 48 nm3
log I(s) 6.70×10-3 6.70×10-4 6.70×10-5 6.70×10-6
Elastin-like polypeptide labeled with unnatural amino acid diBrK small angle scattering data  s, nm-1
ln I(s)
Elastin-like polypeptide labeled with unnatural amino acid diBrK Guinier plot ln 6.70×10-3 Rg: 3.5 nm 0 (3.5 nm)-2 s2
(sRg)2I(s)/I(0)
Elastin-like polypeptide labeled with unnatural amino acid diBrK Kratky plot 1.104 0 3 sRg
p(r)
Elastin-like polypeptide labeled with unnatural amino acid diBrK pair distance distribution function Rg: 3.6 nm 0 Dmax: 14.8 nm

Data validation


There are no models related to this curve.

Synchrotron SAXS data from solutions of Br-labelled ELP in phosphate buffered saline containing 2 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. 2400 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.

Br-labeled ELP in 2 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.

Elastin-like polypeptide labeled with unnatural amino acid diBrK (Br-labeled ELP)
Mol. type   Protein
Organism   synthetic construct
Olig. state   Monomer
Mon. MW   12.9 kDa
Sequence   FASTA