Home Proteases DUB inhibitor - Autophagy activator IU1 For research use only.

IU1

IU1 is a cell-permeable, reversible and selective proteasome inhibitor of human USP14 with IC50 of 4.7 μ M, 25-fold selective to IsoT. IU1 induces autophagy.

IU1 Chemical Structure

IU1 Chemical Structure

CAS No. 314245-33-5

Purity & Quality Control

IU1 Related Products

Signaling Pathway

DUB Signaling Pathways

DUB Signaling Pathway

Cell Data

Cell Lines Assay Type Concentration Incubation Time Formulation Activity Description PMID
HEK293T Function assay 30 uM 2 hrs Inhibition of deubiquitinase in HEK293T cells expressing N-terminal HA-tagged ubiquitin assessed as stabilization of HMW-Ub proteins at 30 uM after 2 hrs by Western blot analysis 30528168
Click to View More Cell Line Experimental Data

Biological Activity

Description IU1 is a cell-permeable, reversible and selective proteasome inhibitor of human USP14 with IC50 of 4.7 μ M, 25-fold selective to IsoT. IU1 induces autophagy.
Targets
USP14 [1]
(cell-free assay)
4.7 μM
In vitro
In vitro IU1 binds specifically to the activated form of USP14. IU1 can potentially inhibit USP14 by preventing its docking on the proteasome, exhibiting little or no activity toward 8 other DUBs, IsoT, UCH37, BAP1, UCH-L1, UCH-L3, USP15, USP2, USP7. USP14 inhibition is rapidly established upon addition of IU1 and rapidly reversed upon its removal. IU1 inhibits USP14 induced chain trimming and decreases electrophoretic mobility of Ub-CCNB species. IU1 enhances proteasomal degradation of Ub-CCNB in the presence of USP14. IU1 promots degradation of tau and depletes TDP-43, ATXN3, and glial fibrillary acidic protein (GFAP) in proteotoxic mechanisms. [1]
Kinase Assay High-throughput screening
Screening is conducted at the ICCB-Longwood screening facility. 10 μL of recombinant USP14 protein are dispensed into each well of a 384-well low volume plate in duplicate, using a Wellmate plate dispenser. 33.3 nL of compound from the library are pin-transferred into the wells using a Seiko pin transfer robotic system, followed by pre-incubation for about 30 min. The last two columns of each plate are used for positive and negative controls for the assay. To initiate the enzyme reaction, 10 μL of VS-proteasome plus Ub-AMC mixture are added to each well, using a Wellmate dispenser. Samples are then incubated for another 45 min. Ub-AMC hydrolysis is measured at Ex355/Em460 using an Envision plate reader. The final concentrations of USP14, VS-proteasome and Ub-AMC are 15 nM, 1 nM and 0.8 μM, respectively. The final concentration of test compound is approximately 17 μM. Enzymes and substrates are prepared in Ub-AMC assay buffer (50 mM Tris-HCl (pH 7.5), 1 mM EDTA, 1 mM ATP, 5 mM MgCl2, 1 mM DTT, and 1 mg/Ml ovalbumin).
Cell Research Cell lines MEF cells
Concentrations ~1.5 mM
Incubation Time 48 hours
Method MTT

Chemical Information & Solubility

Molecular Weight 300.37 Formula

C18H21FN2O
CAS No. 314245-33-5 SDF Download IU1 SDF
Smiles CC1=CC(=C(N1C2=CC=C(C=C2)F)C)C(=O)CN3CCCC3
Storage (From the date of receipt)

In vitro
Batch:

DMSO : 60 mg/mL ( (199.75 mM) Moisture-absorbing DMSO reduces solubility. Please use fresh DMSO.)

Ethanol : 60 mg/mL

Water : Insoluble


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In vivo
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Tech Support

Answers to questions you may have can be found in the inhibitor handling instructions. Topics include how to prepare stock solutions, how to store inhibitors, and issues that need special attention for cell-based assays and animal experiments.

Handling Instructions

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