I3MT-3

Catalog No.S3541 Batch:S354101

Technical Data

Formula	C₁₇H₁₄N₂O₂S
Molecular Weight	310.37	CAS No.	459420-09-8
Solubility (25°C)*	In vitro	DMSO	62 mg/mL (199.76 mM)
		Ethanol	2 mg/mL (6.44 mM)
		Water	Insoluble
* <1 mg/ml means slightly soluble or insoluble. * Please note that Selleck tests the solubility of all compounds in-house, and the actual solubility may differ slightly from published values. This is normal and is due to slight batch-to-batch variations. * Room temperature shipping (Stability testing shows this product can be shipped without any cooling measures.)

Preparing Stock Solutions

Biological Activity

Description

I3MT-3 (HMPSNE, compound 3) is a potent, selective, and cell-membrane permeable inhibitor of 3-mercaptopyruvate sulfurtransferase (3MST) with IC50 of 2.7 μM. I3MT-3 targets a persulfurated cysteine residue located in the active site of 3MST.

Targets

3MST ^[1]
(Cell-free assay)

2.7 μM

In vitro

I3MT-3 (HMPSNE, compound 3), a pharmacological 3-MST inhibitor, exerts a bell-shaped effect on several cellular bioenergetic parameters related to oxidative phosphorylation in CT26 cells, suggesting that the 3-MST/H2S system significantly contributes to the maintenance of cell proliferation, migration and cellular bioenergetics.^[2]

Protocol (from reference)

Cell Assay:^{^[2]}	Cell lines CT26 mouse colon carcinoma cell line Concentrations 1-300 μM Incubation Time 24 h Method The activity of 3-MST in presence of various concentrations of HMPSNE was also tested in CT26 cells homogenates. CT26 cells were centrifuged 5 min at 400× g in order to resuspend the pellet in lysis buffer 150 mM NaCl and 50 mM Tris-HCl, pH 8 containing 1% NP40 and Protease and Phosphatase Inhibitor Cocktail added just before use. Samples were sonicated with ultrasound probe three times a cycle of 5 s on and 5 s off at 70% amplitude and kept on ice for 30 min. The protein concentrations were measured as described above. HMPSNE dilutions were prepared as above and incubated 24 h at 37 °C with 100 µg/well of proteins from CT26 homogenates in black 96-well plate. After treating the homogenate with HMPSNE, 3-mercaptopyruvate was added for a final concentration of 500 µM and AzMC for a final concentration of 10 µM. Fluorescence measurement and analysis were performed.

Cell Assay:^{^[2]}

Cell lines
CT26 mouse colon carcinoma cell line
Concentrations
1-300 μM
Incubation Time
24 h
Method
The activity of 3-MST in presence of various concentrations of HMPSNE was also tested in CT26 cells homogenates. CT26 cells were centrifuged 5 min at 400× g in order to resuspend the pellet in lysis buffer 150 mM NaCl and 50 mM Tris-HCl, pH 8 containing 1% NP40 and Protease and Phosphatase Inhibitor Cocktail added just before use. Samples were sonicated with ultrasound probe three times a cycle of 5 s on and 5 s off at 70% amplitude and kept on ice for 30 min. The protein concentrations were measured as described above. HMPSNE dilutions were prepared as above and incubated 24 h at 37 °C with 100 µg/well of proteins from CT26 homogenates in black 96-well plate. After treating the homogenate with HMPSNE, 3-mercaptopyruvate was added for a final concentration of 500 µM and AzMC for a final concentration of 10 µM. Fluorescence measurement and analysis were performed.

References

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SHIPPING AND STORAGE
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