Design of a Cereblon construct for crystallographic and biophysical studies of protein degraders

Kroupova A, Spiteri V, Rutter Z, Furihata H, Darren D, Ramachandran S, Chakraborti S, Haubrich K, Pethe J, Gonzales D, Wijaya A, Rodriguez-Rios M, Sturbaut M, Lynch D, Farnaby W, Nakasone M, Zollman D, Ciulli A, Nature Communications 15(1) (2024) DOI

SASDVP6 – Cereblon-midi (CRBNmidi), an engineered Cereblon construct for crystallographic and biophysical studies, bound to Boc-AcQ

Cereblon-midi
MWexperimental 39 kDa
MWexpected 37 kDa
VPorod 62 nm3
log I(s) 2.23×10-2 2.23×10-3 2.23×10-4 2.23×10-5
Cereblon-midi small angle scattering data  s, nm-1
ln I(s)
Cereblon-midi Guinier plot ln 2.24×10-2 Rg: 2.4 nm 0 (2.4 nm)-2 s2
(sRg)2I(s)/I(0)
Cereblon-midi Kratky plot 1.104 0 3 sRg
p(r)
Cereblon-midi pair distance distribution function Rg: 2.5 nm 0 Dmax: 9.6 nm

Data validation


There are no models related to this curve.

Synchrotron SAXS data from solutions of CRBNmidi bound to Boc-AcQ in 20 mM HEPES, 500 mM NaCl, 0.5 mM TCEP, pH 7.5 were collected on the B21 beam line at the Diamond Light Source (Didcot, UK) using a Eiger 4M detector at a sample-detector distance of 3.7 m and at a wavelength of λ = 0.09464 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 50.00 μl sample at 3.4 mg/ml was injected at a 0.08 ml/min flow rate onto a Cytiva Superdex 200 Increase 3.2/300 column at 15°C. 600 successive 3 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.

CRBN-midi, an engineered Cereblon construct for crystallographic and biophysical studies of protein degraders (derived from human Cereblon, Uniprot Q96SW2).

Cereblon-midi (CRBNmidi)
Mol. type   Protein
Organism   synthetic construct
Olig. state   Monomer
Mon. MW   37.4 kDa
Sequence   FASTA