| Size | Price | Stock | Qty |
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| 1g |
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| 5g |
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| 10g |
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| Other Sizes |
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Purity: ≥98%
| Targets |
Antiviral; anticancer
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|---|---|
| ln Vitro |
Ribose methylation is among the most ubiquitous modifications found in RNA. 2'-O-methyluridine is found in rRNA, snRNA, snoRNA and tRNA of Archaea, Bacteria, and Eukaryota. Moreover, 2'-O-methylribonucleosides are promising starting materials for the production of nucleic acid-based drugs. Despite the countless possibilities of practical use for the metabolic enzymes associated with methylated nucleosides, there are very few reports regarding the metabolic fate and enzymes involved in the metabolism of 2'-O-alkyl nucleosides. The presented work focuses on the cellular degradation of 2'-O-methyluridine. A novel enzyme was found using a screening strategy that employs Escherichia coli uracil auxotroph and the metagenomic libraries. A 2'-O-methyluridine hydrolase (RK9NH) has been identified together with an aldolase (RK9DPA)-forming a part of a probable gene cluster that is involved in the degradation of 2'-O-methylated nucleosides. The RK9NH is functional in E. coli uracil auxotroph and in vitro. The RK9NH nucleoside hydrolase could be engineered to enzymatically produce 2'-O-methylated nucleosides that are of great demand as raw materials for production of nucleic acid-based drugs. Moreover, RK9NH nucleoside hydrolase converts 5-fluorouridine, 5-fluoro-2'-deoxyuridine and 5-fluoro-2'-O-methyluridine into 5-fluorouracil, which suggests it could be employed in cancer therapy[1].
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| Enzyme Assay |
Substrate Specificity Measurements
The purified recombinant RK9NH nucleoside hydrolase protein was tested for substrate specificity using the thin-layer chromatography (TLC) and high-performance liquid chromatography-mass spectrometry (HPLC-MS) methods. A standard enzymatic reaction of RK9NH was carried out at 37 °C for 1 h and contained 30 mM (uridine, 2′-deoxyuridine, 2′-O-methyluridine, 5-fluorouridine, 5-fluoro-2′-deoxyuridine, 5-fluoro-2′-O-methyluridine) of substrate, final 0.025 mg/mL concentration of recombinant 6×His-tagged RK9NH in 40 µL final volume of 50 mM Tris-HCl pH 8 buffer (the same results were obtained using a 50 mM potassium phosphate buffer pH 7). Not all of the substrates are well soluble in water (and the buffers used), hence some of the final reaction concentrations varied accordingly to the limit of solubility of the substrates: 20 mM cytidine, 2′-deoxycytidine, 5-methyluridine, thymidine; 15 mM 3′-O-methyluridine, 2′-O-allyluridine, 3′-O-allyluridine, 2′-O-methylcytidine, 2′-O-methyladenosine, 2′-O-methylguanosine; 10 mM adenosine, 2′-deoxyadenosine, 2′-amino-2′-deoxyuridine and 5 mM guanosine, 2′-deoxyguanosine, inosine were used. The final 0.017 mg/mL, 0.013 mg/mL, 0.008 mg/mL, 0.004 mg/mL RK9NH protein concentrations were used respectively[1].
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| References | |
| Additional Infomation |
2'-O-methyluridine is a methyluridine that consists of uridine bearing a single methyl substituent located at position O-2' on the ribose ring.
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| Molecular Formula |
C10H14N2O6
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|---|---|
| Molecular Weight |
258.23
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| Exact Mass |
258.085
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| Elemental Analysis |
C, 46.51; H, 5.46; N, 10.85; O, 37.17
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| CAS # |
2140-76-3
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| PubChem CID |
102212
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| Appearance |
Typically exists as white to off-white solids at room temperature
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| Density |
1.5±0.1 g/cm3
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| Melting Point |
154-156ºC
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| Index of Refraction |
1.611
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| LogP |
-1.14
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| Hydrogen Bond Donor Count |
3
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| Hydrogen Bond Acceptor Count |
6
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| Rotatable Bond Count |
3
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| Heavy Atom Count |
18
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| Complexity |
385
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| Defined Atom Stereocenter Count |
4
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| SMILES |
O=C1NC(C=CN1[C@@H]2O[C@H](CO)[C@@H](O)[C@H]2OC)=O
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| InChi Key |
SXUXMRMBWZCMEN-ZOQUXTDFSA-N
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| InChi Code |
InChI=1S/C10H14N2O6/c1-17-8-7(15)5(4-13)18-9(8)12-3-2-6(14)11-10(12)16/h2-3,5,7-9,13,15H,4H2,1H3,(H,11,14,16)/t5-,7-,8-,9-/m1/s1
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| Chemical Name |
C10H14N2O6
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| Synonyms |
O(2')-Methyluridine Uridine, 2'-O-Methyluridine; 2140-76-3; Uridine, 2'-O-methyl-; 2'-O-Methyl Uridine; 1-((2R,3R,4R,5R)-4-hydroxy-5-(hydroxymethyl)-3-methoxytetrahydrofuran-2-yl)pyrimidine-2,4(1H,3H)-dione; O(2')-Methyluridine; 399VZB6TMB; 1-[(2R,3R,4R,5R)-4-hydroxy-5-(hydroxymethyl)-3-methoxyoxolan-2-yl]pyrimidine-2,4-dione; 2'-O-methyl-2'-O-Methyluridine
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| HS Tariff Code |
2934.99.9001
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| Storage |
Powder -20°C 3 years 4°C 2 years In solvent -80°C 6 months -20°C 1 month |
| Shipping Condition |
Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)
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| Solubility (In Vitro) |
DMSO : ~100 mg/mL (~387.25 mM)
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|---|---|
| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.5 mg/mL (9.68 mM) (saturation unknown) in 10% DMSO + 40% PEG300 + 5% Tween80 + 45% Saline (add these co-solvents sequentially from left to right, and one by one), clear solution.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 25.0 mg/mL clear DMSO stock solution to 400 μL PEG300 and mix evenly; then add 50 μL Tween-80 to the above solution and mix evenly; then add 450 μL normal saline to adjust the volume to 1 mL. Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH₂ O to obtain a clear solution. Solubility in Formulation 2: ≥ 2.5 mg/mL (9.68 mM) (saturation unknown) in 10% DMSO + 90% (20% SBE-β-CD in Saline) (add these co-solvents sequentially from left to right, and one by one), clear solution. For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 25.0 mg/mL clear DMSO stock solution to 900 μL of 20% SBE-β-CD physiological saline solution and mix evenly. Preparation of 20% SBE-β-CD in Saline (4°C，1 week): Dissolve 2 g SBE-β-CD in 10 mL saline to obtain a clear solution. View More
Solubility in Formulation 3: ≥ 2.5 mg/mL (9.68 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (add these co-solvents sequentially from left to right, and one by one), clear solution. |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 3.8725 mL | 19.3626 mL | 38.7252 mL | |
| 5 mM | 0.7745 mL | 3.8725 mL | 7.7450 mL | |
| 10 mM | 0.3873 mL | 1.9363 mL | 3.8725 mL |
*Note: Please select an appropriate solvent for the preparation of stock solution based on your experiment needs. For most products, DMSO can be used for preparing stock solutions (e.g. 5 mM, 10 mM, or 20 mM concentration); some products with high aqueous solubility may be dissolved in water directly. Solubility information is available at the above Solubility Data section. Once the stock solution is prepared, aliquot it to routine usage volumes and store at -20°C or -80°C. Avoid repeated freeze and thaw cycles.
Calculation results
Working concentration： mg/mL；
Method for preparing DMSO stock solution： mg drug pre-dissolved in μL DMSO (stock solution concentration mg/mL). Please contact us first if the concentration exceeds the DMSO solubility of the batch of drug.
Method for preparing in vivo formulation:：Take μL DMSO stock solution, next add μL PEG300, mix and clarify, next addμL Tween 80, mix and clarify, next add μL ddH2O，mix and clarify.
(1) Please be sure that the solution is clear before the addition of next solvent. Dissolution methods like vortex, ultrasound or warming and heat may be used to aid dissolving.
(2) Be sure to add the solvent(s) in order.
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