| Size | Price | Stock | Qty |
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| 1mg |
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| 5mg |
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| Other Sizes |
| Targets |
Mannosyltransferase; GTP.
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|---|---|
| ln Vitro |
The α-anomer of GDP-D-mannose is called GDP-α-D-mannose.
GDP-alpha-D-mannose disodium is the essential donor substrate for mannosyltransferase enzymes. It serves as a direct source of mannose residues, which are transferred to growing glycoprotein, glycolipid, and polysaccharide chains during the process of glycosylation. This compound is also the precursor in the biosynthesis of GDP-beta-L-fucose, an important nucleotide sugar used in fucosylation reactions. Furthermore, GDP-alpha-D-mannose has been shown to have inhibitory activities: it acts as a competitive inhibitor against GTP with a Ki value of 14.7 microM, and as a noncompetitive inhibitor against mannose-1-phosphate (mannose-1-P) with a Ki value of 115 microM. These inhibitory properties make it a useful tool for studying the kinetics of enzymes that utilize these substrates, such as mannose-1-phosphate guanylyltransferase. |
| ln Vivo |
GDP-alpha-D-mannose disodium is primarily a research tool for in vitro biochemical experiments. Its role in vivo is a natural part of cellular metabolism, acting as an intermediate in the synthesis of glycoconjugates. While it is not used as a therapeutic drug in vivo, experiments that manipulate its cellular levels (e.g., through genetic modification of its biosynthetic enzymes) have demonstrated its essential role in cell viability, protein folding, and trafficking. In certain disease models involving defective glycosylation (e.g., Congenital Disorders of Glycosylation), the addition of GDP-alpha-D-mannose or its precursors has been explored as a potential rescue strategy.
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| Enzyme Assay |
The enzymatic activity of mannosyltransferases using GDP-alpha-D-mannose as a substrate can be measured using a radioactive or fluorescence-based assay. For a standard assay, a recombinant mannosyltransferase enzyme (e.g., yeast or human PIG-M, an enzyme in the GPI anchor biosynthesis pathway) is expressed and purified. The enzyme is incubated in a suitable reaction buffer (e.g., 50 mM Tris-HCl, pH 7.4, 10 mM MnCl2, 1 mM DTT, and 0.1% Triton X-100) with a range of concentrations of GDP-alpha-D-mannose disodium. The reaction is initiated by the addition of the acceptor substrate (e.g., a synthetic lipid-linked oligosaccharide or a peptide). After incubation at 37degC for a defined period (e.g., 30-60 minutes), the reaction is terminated by boiling or by the addition of a suitable stop solution. If using a radiolabeled donor (e.g., GDP-[3H]-alpha-D-mannose), the radiolabeled product is separated from the unincorporated donor by extraction into an organic solvent or by binding to a filter membrane. The incorporated radioactivity is then measured by scintillation counting. Alternatively, a fluorescence-based assay using a fluorescently labeled acceptor substrate can be used, where the reaction product is resolved by HPLC or electrophoresis and quantified by fluorescence detection. To assess the competitive inhibition of GTP by GDP-alpha-D-mannose, a mannose-1-phosphate guanylyltransferase assay is used. The enzyme is incubated with varying concentrations of mannose-1-P and a fixed concentration of GTP in the presence of different fixed concentrations of GDP-alpha-D-mannose. The formation of GDP-mannose and pyrophosphate is measured. The data is analyzed using a Lineweaver-Burk plot to determine the type of inhibition (competitive vs. noncompetitive) and to calculate the Ki values for both GTP and mannose-1-P.
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| Cell Assay |
In vitro cell culture experiments using GDP-alpha-D-mannose are limited by the fact that nucleotide sugars are not efficiently taken up by intact cells. Therefore, the effects of GDP-alpha-D-mannose in cells are typically studied by manipulating its metabolism rather than adding it directly to the medium. However, in permeabilized cell assays, it can be used. For example, HeLa cells are grown to 80-90% confluency in a T-75 flask. The cells are harvested, washed with ice-cold PBS, and then treated with a permeabilization buffer (e.g., 20 mM HEPES, pH 7.2, 50 mM potassium acetate, 5 mM magnesium acetate, 1 mM EGTA, and 50 ug/mL digitonin) for 10 minutes on ice. The permeabilized cells are then centrifuged and resuspended in an assay buffer. The cells are then incubated with GDP-alpha-D-mannose and a fluorescently labeled acceptor substrate for the mannosyltransferase of interest. After the reaction, the cells are washed, and the product is visualized by fluorescence microscopy or analyzed by flow cytometry.
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| Animal Protocol |
Given that GDP-alpha-D-mannose disodium is an endogenous cellular metabolite and not a xenobiotic drug, there are no standard animal dosing protocols for it as a therapeutic agent. In research settings, it may be administered to animals to study its effects on glycosylation, but it is not typically done due to poor cellular uptake. For studies involving metabolic labeling, its isotopically labeled versions (e.g., ¹3C-labeled) can be administered via intravenous or intraperitoneal injection. For example, mice can be fasted overnight and then injected intraperitoneally with 50 mg/kg of ¹3C-labeled GDP-alpha-D-mannose dissolved in sterile saline. Blood and tissue samples (e.g., liver) are collected at various time points post-injection. The incorporation of the labeled mannose into glycoproteins is analyzed by LC-MS/MS after protein hydrolysis.
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| ADME/Pharmacokinetics |
As an endogenous metabolite, the pharmacokinetics (PK) of GDP-alpha-D-mannose disodium is not typically studied as an administered drug. Its endogenous levels are tightly regulated by cellular metabolism. The cytosolic concentration of GDP-alpha-D-mannose is known to be in the range of 100-200 uM in many cell types. If administered exogenously, it would be rapidly degraded by cellular phosphatases and nucleotidases. Intravenous administration would likely result in a very short plasma half-life, measured in minutes, due to rapid clearance and metabolism. For research purposes, its production and degradation rates can be studied in cells or tissues using stable isotope-labeled precursors.
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| Toxicity/Toxicokinetics |
GDP-alpha-D-mannose disodium is an endogenous cellular metabolite and is considered to have low acute toxicity when used in research settings. It is not classified as a hazardous compound. Its physiological levels are tightly regulated, and dysregulation can lead to disease. For example, mutations in the enzyme that synthesizes it, GDP-mannose pyrophosphorylase (GMPPA or GMPPB), cause congenital disorders of glycosylation (CDG) with severe neuromuscular and developmental abnormalities. Therefore, while the compound itself is not toxic, its depletion is highly detrimental. In vitro, adding very high concentrations (e.g., >5 mM) to cells might lead to osmotic stress, but the compound itself is not known to have specific toxicological mechanisms.
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| References | |
| Additional Infomation |
GDP-alpha-D-mannose is a key intermediate in the biosynthesis of many important glycoconjugates, including N-linked glycoproteins (which are crucial for protein folding and cellular recognition), glycosylphosphatidylinositol (GPI) anchors (which tether proteins to the cell membrane), and plant cell wall polysaccharides. In humans, the synthesis of GDP-alpha-D-mannose from mannose-1-phosphate and GTP is catalyzed by the enzymes GMPPA and GMPPB. The compound is commercially available for research purposes and is often used as a substrate in in vitro assays to determine the activity of mannosyltransferases. It is also used as a biochemical tool to study the mechanisms and specificities of these enzymes, which are potential targets for drug development, particularly for anti-fungal or anti-parasitic agents that target glycosylation pathways essential for pathogen viability but not present in humans.
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| Molecular Formula |
C₁₆H₂₃N₅NA₂O₁₆P₂
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|---|---|
| Molecular Weight |
649.30
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| Exact Mass |
649.041
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| CAS # |
148296-46-2
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| Related CAS # |
GDP-D-mannose disodium;103301-73-1
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| PubChem CID |
135438424
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| Appearance |
White to off-white solid powder
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| Hydrogen Bond Donor Count |
8
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| Hydrogen Bond Acceptor Count |
18
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| Rotatable Bond Count |
9
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| Heavy Atom Count |
41
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| Complexity |
1040
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| Defined Atom Stereocenter Count |
9
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| SMILES |
C1=NC2=C(N1[C@H]3[C@@H]([C@@H]([C@H](O3)COP(=O)([O-])OP(=O)([O-])O[C@@H]4[C@H]([C@H]([C@@H]([C@H](O4)CO)O)O)O)O)O)N=C(NC2=O)N.[Na+].[Na+]
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| InChi Key |
XOAGKSFNHBWACO-RAUZPKMFSA-L
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| InChi Code |
InChI=1S/C16H25N5O16P2.2Na/c17-16-19-12-6(13(28)20-16)18-3-21(12)14-10(26)8(24)5(34-14)2-33-38(29,30)37-39(31,32)36-15-11(27)9(25)7(23)4(1-22)35-15;;/h3-5,7-11,14-15,22-27H,1-2H2,(H,29,30)(H,31,32)(H3,17,19,20,28);;/q;2*+1/p-2/t4-,5-,7-,8-,9+,10-,11+,14-,15-;;/m1../s1
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| Chemical Name |
disodium;[[(2R,3S,4R,5R)-5-(2-amino-6-oxo-1H-purin-9-yl)-3,4-dihydroxyoxolan-2-yl]methoxy-oxidophosphoryl] [(2R,3S,4S,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)oxan-2-yl] phosphate
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| Synonyms |
GDPαDmannose disodium; GDP α D mannose disodium
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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 Note: Please store this product in a sealed and protected environment, avoid exposure to moisture. |
| 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) |
H2O : ~100 mg/mL (~154.01 mM)
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|---|---|
| Solubility (In Vivo) |
Note: Listed below are some common formulations that may be used to formulate products with low water solubility (e.g. < 1 mg/mL), you may test these formulations using a minute amount of products to avoid loss of samples.
Injection Formulations
Injection Formulation 1: DMSO : Tween 80： Saline = 10 : 5 : 85 (i.e. 100 μL DMSO stock solution → 50 μL Tween 80 → 850 μL Saline)(e.g. IP/IV/IM/SC) *Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH ₂ O to obtain a clear solution. Injection Formulation 2: DMSO : PEG300 ：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL DMSO → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline) Injection Formulation 3: DMSO : Corn oil = 10 : 90 (i.e. 100 μL DMSO → 900 μL Corn oil) Example: Take the Injection Formulation 3 (DMSO : Corn oil = 10 : 90) as an example, if 1 mL of 2.5 mg/mL working solution is to be prepared, you can take 100 μL 25 mg/mL DMSO stock solution and add to 900 μL corn oil, mix well to obtain a clear or suspension solution (2.5 mg/mL, ready for use in animals). View More
Injection Formulation 4: DMSO : 20% SBE-β-CD in saline = 10 : 90 [i.e. 100 μL DMSO → 900 μL (20% SBE-β-CD in saline)] Oral Formulations
Oral Formulation 1: Suspend in 0.5% CMC Na (carboxymethylcellulose sodium) Oral Formulation 2: Suspend in 0.5% Carboxymethyl cellulose Example: Take the Oral Formulation 1 (Suspend in 0.5% CMC Na) as an example, if 100 mL of 2.5 mg/mL working solution is to be prepared, you can first prepare 0.5% CMC Na solution by measuring 0.5 g CMC Na and dissolve it in 100 mL ddH2O to obtain a clear solution; then add 250 mg of the product to 100 mL 0.5% CMC Na solution, to make the suspension solution (2.5 mg/mL, ready for use in animals). View More
Oral Formulation 3: Dissolved in PEG400 (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 1.5401 mL | 7.7006 mL | 15.4012 mL | |
| 5 mM | 0.3080 mL | 1.5401 mL | 3.0802 mL | |
| 10 mM | 0.1540 mL | 0.7701 mL | 1.5401 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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