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 |