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Chemical Properties Biological Description Data Sheet (Cat.No.T2154) MG132 Chemical Properties CAS No.: 133407-82-6 Formula： C26H41N3O5 Molecular Weight： 475.62 Appearance： N/A Storage： 0-4℃ for short term (days to weeks), or -20℃ for long term (months). Biological Description Description MG-132 is a potent cell-permeable 20S proteasome inhibitor (IC50: 100 nM). It also inhibits calpain (IC50: 1.2 μM). Targets(IC50) Proteasome: 100nM In vitro The IC50 of MG-132 for the alkali-denatured casein-degrading activity of m-calpain was around 1.2 μM. The IC50 for the ZLLL MCA-degrading activity of 20S proteasome by MG-132 was 100 nM [1]. It also inhibits proteasomal chymotrypsin-like peptidase activity (IC50: 24.2 nM) [2]. Dose-dependent inhibition of cell growth was observed in HeLa cells with an IC50 of approximately 5 μM MG132 for 24 h. The treatment with MG132 induced S, G2-M or non-specific phase arrests of the cell cycle dose-dependently. Treatment with MG132 induced apoptosis in a dose-dependent manner, as evidenced by sub-G1 cells and annexin V staining cells [3]. In vivo MG132 (0.1 mg/kg/day) was intraperitoneally injected to rats with abdominal aortic banding (AAB) for 8 weeks. Results showed that treatment with MG132 significantly attenuated left ventricular (LV) myocyte area, LV weight/body weight, and lung weight/body weight ratios, decreased LV diastolic diameter and wall thickness and increased fractional shortening in AAB rats. AAB induced the phosphorylation of ERK1/2, JNK1, and p38 in cardiac myocytes. The elevated phosphorylation levels of ERK1/2 and JNK1 in AAB rats were significantly reversed by MG132 treatment [4]. MG132 significantly inhibited IκBα degradation thus preventing NFκB activation in vitro. MG132 preserved muscle and myofiber cross-sectional area by downregulating the muscle- specific ubiquitin ligases atrogin-1/MAFbx and MuRF-1 mRNA in vivo. This effect resulted in a diminished rehabilitation period [5]. Kinase Assay Inhibitory activities of ZLLa1 and ZLLLal against m-calpain and 20S proteasome were measured by previously described methods.For the m-calpain inhibitory assay,the 0.5 ml reaction mixture contained 0.24% alkali- denatured casein,28 mM 2-mercaptoethanol,0.94 unit of m-calpain,ZLLal or ZLLLal,6 mM CaCl2,and 0.1M Tris- HC1 (pH 7.5).The reaction was started by the addition of m-calpain solution and stopped by the addition of 0.5 ml of 10% trichloroacetic acid after incubation at 30℃ for 15 min.After centrifugation at 1,300×g for 10 min,the absorbance of the supernatant at 280 nm was measured.The reaction mixture for the 20S proteasome inhibitory assay contained 0.1 M Tris-acetate,pH 7.0,20S proteasome,ZLLa1 or ZLLLal,and 25 μM substrate dissolved in dimethyl sulfoxide in a final volume of 1 ml.After incubation at 37℃ for 15 min,the reaction was stopped by the addition of 0.1 ml of 10% SDS and 0.9 ml of 0.1 M Tris-acetate,pH 9.0.The fluorescence of the reaction products was measured.To determine the IC50s against m-calpain and 20S proteasome,various concentrations of the synthetic peptide aldehydes were included in the assay mixture [1]. Page 1 of 3 www.targetmol.com A DRUG SCREENING EXPERT Cell Research Cells are exposed to various concentrations of MG-132 for 24, and 48 hours. Supernatant and monolayer cells are harvested by centrifugation and fixed in 70% ethanol in PBS for staining with acridine orange. Equal volumes of cells and acridine orange (5 mg/mL in PBS) are mixed on a microscope slide and examined by fluorescence microscopy. For annexin V analysis, cells are harvested by centrifugation and stained with annexin V and propidium iodide. For cell cycle analysis, cells are rehydrated in PBS at room temperature for 10 minutes, followed by staining with propidium iodide (5 mg/mL). All samples are analyzed using a Coulter Epics XL flow cytometer. (Only for Reference) Cell lines：KIM-2, HC11, and ES Animal Animal Model：mdx (C57BL/10ScSn DMD mdx) mice Research Solubility Information Solubility Ethanol: 47.5 mg/mL (100 mM) DMSO: 90 mg/mL (189.23 mM) Water: Insoluble (< 1 mg/ml refers to the product slightly soluble or insoluble) Preparing Stock Solutions 1mg 5mg 10mg 1 mM 2.103 mL 10.513 mL 21.025 mL 5 mM 0.421 mL 2.103 mL 4.205 mL 10 mM 0.210 mL 1.051 mL 2.103 mL 50 mM 0.042 mL 0.210 mL 0.421 mL Please select the appropriate solvent to prepare the stock solution, according to the solubility of the product in different solvents. The storage conditions and period of the stock solution: - 80 ℃ for 6 months; - 20 ℃ for 1 month. Please use it as soon as possible. Reference 1. Tsubuki S, et al. Differential inhibition of calpain and proteasome activities by peptidyl aldehydes of di-leucine and tri-leucine. J Biochem. 1996 Mar;119(3):572-6. 2. Braun HA, et al. Tripeptide mimetics inhibit the 20 S proteasome by covalent bonding to the active threonines. J Biol Chem. 2005 Aug 5;280(31):28394-401. 3. Han YH, et al. The effect of MG132, a proteasome inhibitor on HeLa cells in relation to cell growth, reactive oxygen species and GSH. Oncol Rep. 2009 Jul;22(1):215-21. 4. Chen B, et al. MG132, a proteasome inhibitor, attenuates pressure-overload-induced cardiac hypertrophy in rats by modulation of mitogen-activated protein kinase signals. Acta Biochim Biophys Sin (Shanghai). 2010 Apr;42(4):253-8. 5. Caron AZ, et al. The proteasome inhibitor MG132 reduces immobilization-induced skeletal muscle atrophy in mice. BMC Musculoskelet Disord. 2011 Aug 15;12:185. 6. Matsumoto Y, et al. Enhanced efficacy against cervical carcinomas through polymeric micelles physically incorporating theproteasome inhibitor MG132. Cancer Sci. 2016 Jun;107(6):773-81. 7. Harhouri K, et al. MG132-induced progerin clearance is mediated by autophagy activation and splicing regulation. EMBO Mol Med. 2017 Sep;9(9):1294-1313. 8. Guzmán-Téllez P, Martínez-Valencia D, Silva-Olivares A, et al. Naegleria fowleri and Naegleria gruberi 20S proteasome: identification and characterization[J]. European Journal of Cell Biology. 2020: 151085 9. Wu H, Cheng J, Huang Y, et al. TRIM21 and PHLDA3 Negatively Regulate the Cross-Talk between the PI3K/AKT Pathway and PPP Metabolism. Nature Communications. 2020, 11(1): 1-16. 10. Luo Q, Wu X, Zhao P, et al. OTUD1 Activates Caspase‐Independent and Caspase‐Dependent Apoptosis by Promoting AIF Nuclear Translocation and MCL1 Degradation[J]. Advanced Science. 2021: 2002874. Page 2 of 3 www.targetmol.com A DRUG SCREENING EXPERT Inhibitors · Natural Compounds · Compound Libraries This product is for Research Use Only · Not for Human or Veterinary or Therapeutic Use. Tel:781-999-4286 E-mail:[email protected] Address:36 Washington Street,Wellesley Hills,MA 02481 Page 3 of 3 www.targetmol.com.
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