@article{8ff1892aa1d841018ace1c07c8ab961b,
title = "Structural Basis and Kinetic Pathway of RBM39 Recruitment to DCAF15 by a Sulfonamide Molecular Glue E7820",
abstract = "E7820 and indisulam are two examples of aryl sulfonamides that recruit RBM39 to Rbx-Cul4-DDA1-DDB1-DCAF15 E3 ligase complex, leading to its ubiquitination and degradation by the proteasome. To understand their mechanism of action, we performed kinetic analysis on the recruitment of RBM39 to DCAF15 and solved a crystal structure of DDA1-DDB1-DCAF15 in complex with E7820 and the RRM2 domain of RBM39. E7820 packs in a shallow pocket on the surface of DCAF15 and the resulting modified interface binds RBM39 through the α1 helix of the RRM2 domain. Our kinetic studies revealed that aryl sulfonamide and RBM39 bind to DCAF15 in a synergistic manner. The structural and kinetic studies confirm aryl sulfonamides as molecular glues in the recruitment of RBM39 and provide a framework for future efforts to utilize DCAF15 to degrade other proteins of interest.",
keywords = "DCAF15, E7820, PROTAC, RBM39, indisulam, molecular glue, sulfonamide, targeted protein degradation",
author = "Xinlin Du and Volkov, {Oleg A.} and Czerwinski, {Robert M.} and Tan, {Hui Ling} and Carlos Huerta and Morton, {Emily R.} and Rizzi, {Jim P.} and Wehn, {Paul M.} and Rui Xu and Deepak Nijhawan and Wallace, {Eli M.}",
note = "Funding Information: This research was funded by Peloton Therapeutics, which was later acquired by Merck. X-ray diffraction data were collected at the Advanced Light Source, which is a Department of Energy (DOE) Office of Science User Facility under contract no. DE-AC02-05CH11231 and Advanced Photon Source, a DOE Office of Science User Facility operated by Argonne National Laboratory under contract no. DE-AC02-06CH11357. X.D. designed and made constructs for recombinant protein expression. O.A.V. and H.T. prepared the viruses for DDA1, DDB1, DDB1?B, DCAF15, and DCAF15?pro. H.T. worked out the procedure for the purification of DDB1?B-DCAF15?pro complex. O.A.V. purified the DDA1-DDB1-DCAF15?pro complex for crystallization and the DDA1-DDB1?B-DCAF15?pro for biophysical assays. X.D. purified R1R2 and RRM2, prepared the tetraplexes, and set up screens for crystallization. X.D. C.H. and O.A.V. optimized crystallization conditions. X.D. collected and processed the data. X.D. and O.A.V. solved the structure and built the models. O.A.V. refined the structure. R.M.C. and E.R.M. designed and performed the FRET and BLI experiments. X.D. carried out the ITC experiments. P.M.W. and R.X. synthesized PT7795. X.D. wrote the manuscript. X.D. J.P.R. and E.M.W. oversaw the project. The authors declare no competing interests. Funding Information: This research was funded by Peloton Therapeutics , which was later acquired by Merck. X-ray diffraction data were collected at the Advanced Light Source, which is a Department of Energy (DOE) Office of Science User Facility under contract no. DE-AC02-05CH11231 and Advanced Photon Source, a DOE Office of Science User Facility operated by Argonne National Laboratory under contract no. DE-AC02-06CH11357. Publisher Copyright: {\textcopyright} 2019 Elsevier Ltd",
year = "2019",
month = nov,
day = "5",
doi = "10.1016/j.str.2019.10.005",
language = "English (US)",
volume = "27",
pages = "1625--1633.e3",
journal = "Structure with Folding & design",
issn = "0969-2126",
publisher = "Cell Press",
number = "11",
}