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Assay Detail

CHEMBL5732769

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Binding
Ubiquitin Activating Enzyme (UAE) HTRF Assay: The UAE enzymatic reaction totals 50 μL and contains 50 mmol HEPES (pH 7.5), 0.05% BSA, 2.5 mM MgCl2, 0.1 uM ATP, 8 nM GST-Ubc-2, 35 nM flag-ubiquitin, 1 nM recombinant human UAE or mouse UAE. Compounds for this hUAE IC50 assay are tested at 10 point 3-fold dilution. The top concentration for this assay is 1 μM. Each compound is ordered in duplicate on the same plate. The enzymatic reaction mixture is incubated for 90 minutes at room temperature (24 degrees C.) in a 384 well plate prior to termination with a stop solution (0.1 M HEPES/0.05% Tween20, 20 mmol EDTA, 410 mM KF, 0.5 nM Eu Cryptate anti-FLAG M2-K antibody (Cis-bio International), 8 ug/mL Anti-GST XL-APC (Prozyme)). After incubation for 120 minutes, quantification of the FRET is performed on the Pherostar (BMG).
375
Total Activities
120
Compounds Tested
3
Activity Types
0
Assay Parameters

Assay Information

Assay Type Binding
Organism Homo sapiens
Confidence 9 — Direct single protein target
Curated By Autocuration

Target

Ubiquitin-like modifier-activating enzyme 1 (CHEMBL5924)
Type SINGLE PROTEIN
Organism Homo sapiens

Publication

Pyrazolopyrimidinyl inhibitors of ubiquitin-activating enzyme
(2019)

Activity Statistics

Type Count Avg pChEMBL Best pChEMBL
IC50 125 6.93 7.26
kon 125 - -
k_off 125 - -

Compounds Tested

Compound Name Phase Activities Best pChEMBL
CHEMBL5864078 3 7.26
CHEMBL6053122 3 7.26
CHEMBL6065415 3 7.26
CHEMBL5997651 3 7.26
CHEMBL5828810 3 7.26
CHEMBL5832385 3 7.26
CHEMBL5971124 3 7.26
CHEMBL5747752 3 7.26
CHEMBL5940945 3 7.26
CHEMBL5918592 3 7.26
CHEMBL5954380 3 7.26
CHEMBL5803994 3 7.26
CHEMBL6003529 3 7.26
CHEMBL5768102 3 7.26
CHEMBL5887314 3 7.26
CHEMBL5780833 3 7.26
CHEMBL5759269 3 7.26
CHEMBL5958476 3 7.26
CHEMBL5743820 3 6.26
CHEMBL5769060 3 6.26
CHEMBL5960054 3 6.26
CHEMBL5929395 3 6.26
CHEMBL5762899 3 6.26
CHEMBL6054356 3 6.26
CHEMBL5799754 3 6.26
CHEMBL5866556 3 6.26
CHEMBL5793677 3 6.26
CHEMBL6020119 6 -
CHEMBL5917491 3 -
CHEMBL6012674 3 -

Activity Data

Compound Name Type Rel. Value Units pChEMBL
CHEMBL5887314 IC50 = 55.0 nM 7.26
CHEMBL6065415 IC50 = 55.0 nM 7.26
CHEMBL5997651 IC50 = 55.0 nM 7.26
CHEMBL5971124 IC50 = 55.0 nM 7.26
CHEMBL5780833 IC50 = 55.0 nM 7.26
CHEMBL5747752 IC50 = 55.0 nM 7.26
CHEMBL5864078 IC50 = 55.0 nM 7.26
CHEMBL6053122 IC50 = 55.0 nM 7.26
CHEMBL5803994 IC50 = 55.0 nM 7.26
CHEMBL5954380 IC50 = 55.0 nM 7.26
CHEMBL5768102 IC50 = 55.0 nM 7.26
CHEMBL5832385 IC50 = 55.0 nM 7.26
CHEMBL6003529 IC50 = 55.0 nM 7.26
CHEMBL5940945 IC50 = 55.0 nM 7.26
CHEMBL5828810 IC50 = 55.0 nM 7.26
CHEMBL5759269 IC50 = 55.0 nM 7.26
CHEMBL5958476 IC50 = 55.0 nM 7.26
CHEMBL5918592 IC50 = 55.0 nM 7.26
CHEMBL5762899 IC50 = 550.0 nM 6.26
CHEMBL5929395 IC50 = 550.0 nM 6.26
CHEMBL6054356 IC50 = 550.0 nM 6.26
CHEMBL5799754 IC50 = 550.0 nM 6.26
CHEMBL5743820 IC50 = 550.0 nM 6.26
CHEMBL5793677 IC50 = 550.0 nM 6.26
CHEMBL5960054 IC50 = 550.0 nM 6.26
CHEMBL5769060 IC50 = 550.0 nM 6.26
CHEMBL5866556 IC50 = 550.0 nM 6.26
CHEMBL5781268 k_off = - s-1 -
CHEMBL5945158 k_off = - s-1 -
CHEMBL5945158 kon = - -
CHEMBL5945158 IC50 < 10.0 nM -
CHEMBL5983436 k_off = - s-1 -
CHEMBL5983436 kon = - -
CHEMBL5781268 IC50 < 10.0 nM -
CHEMBL5913390 k_off = - s-1 -
CHEMBL5913390 kon = - -
CHEMBL5913390 IC50 < 10.0 nM -
CHEMBL5870046 k_off = - s-1 -
CHEMBL5870046 kon = - -
CHEMBL5870046 IC50 < 10.0 nM -
CHEMBL5811200 k_off = - s-1 -
CHEMBL5811200 kon = - -
CHEMBL5811200 IC50 < 10.0 nM -
CHEMBL5873067 k_off = - s-1 -
CHEMBL5873067 kon = - -
CHEMBL5873067 IC50 < 10.0 nM -
CHEMBL5924753 k_off = - s-1 -
CHEMBL5924753 kon = - -
CHEMBL5835205 kon = - -
CHEMBL5924753 IC50 < 10.0 nM -