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. Author manuscript; available in PMC: 2024 Mar 2.
Published in final edited form as: Mol Cell. 2023 Jan 23;83(5):731–745.e4. doi: 10.1016/j.molcel.2022.12.033

KEY RESOURCES TABLE.

REAGENT or RESOURCE SOURCE IDENTIFIER
Antibodies
Rabbit monoclonal anti-GAPDH antibody Cell Signaling Cat# 2118; RRID:AB_561053
Mouse monoclonal anti-Flag antibody Sigma Cat# F1804; RRID:AB_262044
Mouse monoclonal anti-Flag antibody Cell Signaling Cat# 2276; RRID:AB_331783
Bacterial and Virus Strains
BL21-RIPL cells BL21-RIPL cells Cat# 230280
Chemicals, Peptides, and Recombinant Proteins
Ubiquitin Boston Biochem Cat# U-100H-10M
MG132 Selleck Chemicals Cat# S2619
cycloheximide Cell Signaling Cat# 2112
Effectene Transfection Reagent Qiagen Cat# 301425
Imidazole Sigma Cat# I2399
Deposited Data
Cryo-EM structure of wild-type SPOP This study PDB: 8DWV; EMDB: EMD-27761
Cryo-EM structure of E47K SPOP This study PDB: 8DWS; EMDB: EMD-27758
Cryo-EM structure of W22R SPOP form 1 This study PDB: 8DWU; EMDB: EMD-27760
Cryo-EM structure of W22R SPOP form 2 This study PDB: 8DWT; EMDB: EMD-265983
Other data This study Mendeley: DOI: 10.17632/kn52rzcckn.1
Experimental Models: Cell Lines
Human: T-REx-293 Thermo Fisher Scientific Cat# R71007
Recombinant DNA
His-SUMO-SPOP1-374 Marzahn et al.45 N/A
His-SUMO-SPOP1-374 W22R This study N/A
His-SUMO-SPOP1-374 E47K This study N/A
His-SUMO-SPOP1-374 E78K This study N/A
His-BRD3 pET28a This study N/A
pET-DUET-1-His-Cul3/Rbx1 Small et al.46 N/A
GST pGEX4T1 GST-ThrombinNEDD8 Walden et al.47 N/A
pGEX4T1 GST-Thrombin- UBCH5 Huang et al.48 N/A
pGEX4T1 GST-Thrombin-UBC12 Duda et al.40 N/A
pcDNA4/TO-SPOP-Myc wild-type This study N/A
pcDNA4/TO-SPOP-Myc W131G This study N/A
pcDNA4/TO-SPOP-Myc W22R This study N/A
pcDNA4/TO-SPOP-Myc E78K This study N/A
pcDNA4/TO-SPOP-Myc E47K This study N/A
pcDNA3-Myc-Cul3 Ohta et al.49 Addgene #19893
pcDNA3-HA-Rbx1 Ohta et al.49 Addgene #19897
Ub-His Wang et al.50
BRD3-Flag Wai et al.51
His-SUMO-SPOP1-374 W131G This study N/A
Software and Algorithms
cryoSPARC Pujani et al.52 https://cryosparc.com/
Coot Emsley et al.53 http://www2.mrc-lmb.cam.ac.uk/personal/pemsley/coot
Phenix Afonine et al.54 https://www.phenix-online.org
MolProbity Chen et al.55 http://molprobity.biochem.duke.edu
Chimera Pettersen at al.56 https://www.cgl.ucsf.edu/chimera
PyMOL Schrodinger http://www.pymol.org
Other
C-flat Holey Carbon Grid Gold 1.2 μm hole 1.3 μm space 400 mesh Electron Microscopy Sciences Cat# CF413-50-Au
C-flat Holey Carbon Grid 2.0 μm hole 2.0 μm space 300 Mesh Electron Microscopy Sciences Cat# CF-223C-50