Synchrotron SAXS
data from solutions of
Cereblon-midi (CRBNmidi), an engineered Cereblon construct for crystallographic and biophysical studies, bound to Mezigdomide
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 storage ring
(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 6.5 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.
SEC-SAXS of CRBN-midi in complex with mezigdomide (PubChem: https://pubchem.ncbi.nlm.nih.gov/compound/137379043, ChEMBL: https://www.ebi.ac.uk/chembl/compound_report_card/CHEMBL4648616/).
Before SEC, CRBN-midi (6.5 mg/ml, 175 µM) were incubated with 400 µM mezigdomide in SEC buffer with 2 % DMSO.
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