Mln7243

Manufactured by Chemietek

MLN7243 is a chemical compound that functions as a small molecule inhibitor. It is designed for laboratory research purposes.

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Lab products found in correlation

Cycloheximide (chx), by Merck Group (5 mentions) Bortezomib, by Selleck Chemicals (4 mentions) Mg132, by Selleck Chemicals (4 mentions) Mln4924, by MedChemExpress (3 mentions) Bafilomycin a1, by Merck Group (3 mentions) Lipofectamine rnaimax, by Thermo Fisher Scientific (2 mentions) Mln4924, by Selleck Chemicals (2 mentions) Elacridar, by Selleck Chemicals (2 mentions) Mitoxantrone, by Merck Group (2 mentions) Tariquidar, by MedChemExpress (2 mentions) Pf 3758309, by Apexbio (2 mentions) Gedatolisib, by Apexbio (2 mentions) Ks176, by MedChemExpress (2 mentions) Mln4924, by Chemietek (2 mentions) Thz531, by Cayman Chemical (2 mentions) Cudc 101, by MedChemExpress (2 mentions) Ko143, by Cayman Chemical (2 mentions) Anisomycin, by Merck Group (2 mentions) Berzosertib, by Selleck Chemicals (2 mentions) Doxycycline, by Thermo Fisher Scientific (2 mentions)

Spelling variants of this product

MLN7243 (CT-M7243) (5 citations) TAK-243 (MLN7243) (3 citations)

13 protocols using mln7243

Proteasome Regulation and NEDD8 Identification

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Most common chemicals were purchased from Sigma Aldrich. MLN4924 (Takeda Pharmaceuticals), MLN7243 (Chemietek), MG132 (Viva Bioscience), Lipofectamine RNAiMAX (Invitrogen), siRNA On-TARGETplus SMARTpools (Dharmacon), protease Inhibitor Cocktail Tablets EDTA-free, Fugene6 HD (Roche), Suc-LLVY-AMC peptide (BostonBiochem). Rabbit monoclonal anti-NEDD8 (1:2000), Y297 (GeneTex, GTX61205), FK2 mouse anti-ubiquitin, stainings (1:250) (Viva Bioscience, VB2500), rabbit anti-ubiquitin (1:2000), western blotting (DAKO, Z0458), mouse anti-fibrilarin (1:1000) (ab4566), rabbit anti-nucleolin (1:1000) (ab22758), mouse anti-GAPDH (1:5000) (6C5, ab8245), rabbit anti-RPL7 (1:2000) (ab72550) (Abcam), mouse anti-tubulin (1:2000) (Cell Signalling, 3873), mouse anti-HA (1:2000) (12C5, 11583816001), mouse anti-GFP (1:500) (11814460001) (Roche), mouse anti-a6 proteasome subunit (1 μg/ml) (Enzo Life Sciences, BML-PW8100), rabbit polyclonal anti-HUWE1 (1:2000) (Bethyl laboratories, A300-486A), mouse monoclonal anti-p21 (1 μg/ml) (F-5, sc-6246, Santa Cruz), rabbit polyclonal anti-CDT1 (1:1000) (# 06-1295, Millipore), goat anti-mouse Alexa Fluor® 488 (115-545-146), goat Anti-Rabbit Alexa Fluor® 594 (111-585-008) (Jackson ImmunoResearch).

Maghames C.M., Lobato-Gil S., Perrin A., Trauchessec H., Rodriguez M.S., Urbach S., Marin P, & Xirodimas D.P. (2018). NEDDylation promotes nuclear protein aggregation and protects the Ubiquitin Proteasome System upon proteotoxic stress. Nature Communications, 9, 4376.

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Cell Viability Assay with MLN4924

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Cell lines were treated with different concentrations of MLN4924 (Selleck Chemicals), MLN7243 (Chemie Tek), bortezomib (Selleck Chemicals), or a DMSO vehicle control in technical duplicates. After 3 days, viability assays were performed using PrestoBlue (Invitrogen) and a Tecan plate reader. After subtracting the media blank, viability was calculated following normalization to the DMSO vehicle control. Values represent an average of 3 independent experiments.

McGrail D.J., Garnett J., Yin J., Dai H., Shih D.J., Lam T.N., Li Y., Sun C., Li Y., Schmandt R., Wu J.Y., Hu L., Liang Y., Peng G., Jonasch E., Menter D., Yates M.S., Kopetz S., Lu K., Broaddus R., Mills G.B., Sahni N, & Lin S.Y. (2020). Proteome instability is a therapeutic vulnerability in mismatch repair deficient cancer. Cancer cell, 37(3), 371-386.e12.

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Protein Synthesis Inhibition Assay

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Cells were evenly divided into 6- or 12-well plates and transfected with designated plasmids. Twenty four to forty eight hours after transfection, the cells were then treated with the protein synthesis inhibitor CHX (Cayman Chemical, 40–50 μg/ml) for the indicated times before collection. For the inhibitors, MG-132 (Fisher Scientific), bafilomycin (Sigma-Aldrich), and MLN7243 (Chemietek) were added at the same time as CHX.

Tsai M., Osman W., Adair J., ElMergawy R., Chafin L., Johns F., Farkas D., Elhance A., Londino J, & Mallampalli R.K. (2022). The E3 ligase subunit FBXO45 binds the interferon-λ receptor and promotes its degradation during influenza virus infection. The Journal of Biological Chemistry, 298(12), 102698.

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Investigating FBXO Protein Functions

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The following drugs and small molecules were used: MLN7243 (CT-M7243, Chemietek); Leu-Leu methyl ester hydrobromide (L7393, MilliporeSigma); CHX (C7698, MilliporeSigma); E-64d (13533, Cayman); 2-hydroxypropyl-β-cyclodextrin (H-107, MilliporeSigma); vinblastine sulfate salt (catalog V1377, MilliporeSigma); filipin (F9765, MilliporeSigma). Plasmid encoding HA-FBXO2 was a gift from Henry Paulson (University of Michigan, Ann Arbor, Michigan, USA). Plasmids encoding FLAG-FBXO2, FLAG-FBXO6, and FLAG-FBXO27 were gifts from Yukiko Yoshida (Tokyo Metropolitan Institute of Medical Science, Tokyo, Japan). Plasmid encoding EGFP-hGal3 was from addgene (73080).

Liu E.A., Schultz M.L., Mochida C., Chung C., Paulson H.L, & Lieberman A.P. (2020). Fbxo2 mediates clearance of damaged lysosomes and modifies neurodegeneration in the Niemann-Pick C brain. JCI Insight, 5(20), e136676.

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Evaluating Inhibitors of ABC Transporters

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MLN7243 was provided by ChemieTek (Indianapolis, IN). Fumitremorgin C (FTC) was a gift from Dr. Susan Bates (Columbia University, NY). All the drug stock solutions were dissolved in DMSO. The reagents were purchased from Sigma Chemical Co (St. Louis, MO) unless stated otherwise.

Wu Z.X., Yang Y., Wang J.Q., Narayanan S., Lei Z.N., Teng Q.X., Zeng L, & Chen Z.S. (2021). Overexpression of ABCG2 Confers Resistance to MLN7243, a Ubiquitin-Activating Enzyme (UAE) Inhibitor. Frontiers in Cell and Developmental Biology, 9, 697927.

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USP7 Inhibitor Protocols Synthesis

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P5091, MG132 and Bortezomib were purchased from SelleckChem. P22077 and HBX41108 were obtained from TOCRIS. MLN7243 was purchased from Chemietek. Bafilomycin A1 was obtained from MilliporeSigma. The rest of known USP7 inhibitors, WNT974, and TNKS656 were synthesized according to published literature procedures by Novartis.

Ji L., Lu B., Zamponi R., Charlat O., Aversa R., Yang Z., Sigoillot F., Zhu X., Hu T., Reece-Hoyes J.S., Russ C., Michaud G., Tchorz J.S., Jiang X, & Cong F. (2019). USP7 inhibits Wnt/β-catenin signaling through promoting stabilization of Axin. Nature Communications, 10, 4184.

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Comprehensive Compound Acquisition Protocol

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Carfilzomib, MLN4924, and CB-5083 were purchased from Selleck Chemicals. MLN7243 was purchased from Chemie Tek. Cycloheximide, E64, and ALLM were purchased from Sigma-Aldrich. AAF-CMK was purchased from Enzo Life Sciences. Brefeldin A and leupeptin were purchased from Alfa Aesar. Cisplatin was purchased from R&D Systems. CSN5i-3 and ONX-0914 were purchased from Thermo Fisher Scientific. PR-825 and the ATG7 inhibitor, compound #37 (21 (link)), were synthesized by WuXi AppTec.

Mamrosh J.L., Sherman D.J., Cohen J.R., Johnston J.A., Joubert M.K., Li J., Lipford J.R., Lomenick B., Moradian A., Prabhu S., Sweredoski M.J., Vander Lugt B., Verma R, & Deshaies R.J. (2023). Quantitative measurement of the requirement of diverse protein degradation pathways in MHC class I peptide presentation. Science Advances, 9(25), eade7890.

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Detailed Compound Sourcing Protocol

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Mitoxantrone was obtained from Sigma-Aldrich (St. Louis, MO). Pheophorbide a was from Frontier Specialty Chemicals (Logan, UT). MLN-7243 and MLN-4924 were purchased from ChemieTek (Indianapolis, IN). THZ531 and Ko143 were obtained from Cayman Chemical (AnnArbor, MI). PF-3758309 and Gedatolisib were from ApexBio Technology (Houston, TX). CUDC-101, KS176 and tariquidar were from MedChem Express (Monmouth Junction, NJ). Elacridar was from Selleck chemicals (Houston, TX).

Thomas J.R., Frye W.J., Robey R.W., Warner A.C., Butcher D., Matta J.L., Morgan T.C., Edmondson E.F., Salazar P.B., Ambudkar S.V, & Gottesman M.M. (2024). Abcg2a is the functional homolog of human ABCG2 expressed at the zebrafish blood–brain barrier. Fluids and Barriers of the CNS, 21, 27.

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Cellular Fractionation and Protein Inhibition

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P22077 (Merck-Millipore), NMS873 (Tocris), cycloheximide (Sigma, Merck), ML792 (Synthetized in the CNIO) and MLN7243 (Chemietek) were dissolved in DMSO; cells were incubated for the indicated time in the presence of the inhibitor or an equivalent amount of DMSO. Whole cell extracts were prepared by lysing cells in 50 mM Tris, pH 7.5, 8 M Urea, and 1% Chaps. Cytosolic and nuclear extracts were prepared following the protocol described before (Lecona et al., 2008 (link)) and the chromatin fraction was then extracted in 50 mM Tris, pH 7.5, 8 M Urea, and 1% Chaps. Transfection of RPE cells with specific siRNA was carried out using Lipofectamine RNAimax (Invitrogen, Thermo Fisher Scientific) according to the manufacturer’s instructions and using pools of 4 specific siRNA directed against the indicated proteins (Dharmacon, Horizon Discovery). Transfection of HCT116 cells with the pCL-His-hUbi plasmid (Young et al., 2011 (link)) was carried out using Lipofectamine 2000 (Invitrogen, Thermo Fisher Scientific) following the manufacturer’s instructions.

Franz A., Valledor P., Ubieto-Capella P., Pilger D., Galarreta A., Lafarga V., Fernández-Llorente A., de la Vega-Barranco G., den Brave F., Hoppe T., Fernandez-Capetillo O, & Lecona E. (2021). USP7 and VCPFAF1 define the SUMO/Ubiquitin landscape at the DNA replication fork. Cell Reports, 37(2), 109819.

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Acquisition of Diverse Chemical Compounds

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Thomas J.R., Frye W.J., Robey R.W., Warner A.C., Butcher D., Matta J.L., Morgan T.C., Edmondson E.F, & Gottesman M.M. (2023). Abcg2a is the functional homolog of human ABCG2 expressed at the zebrafish blood-brain barrier. bioRxiv.

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