| Size | Price | Stock | Qty |
|---|---|---|---|
| 100mg |
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| 500mg | |||
| 1g | |||
| Other Sizes |
| Targets |
1. Galactose 1-phosphate uridylyltransferase (GALT, the enzyme for which Galactose 1-phosphate Potassium salt acts as a natural substrate, as it is a substrate rather than an inhibitor of GALT) [1]
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|---|---|
| ln Vitro |
1. Substrate for GALT enzymatic catalysis: Galactose 1-phosphate Potassium salt serves as the key substrate for recombinant wild-type human GALT in vitro. In the presence of UDP-glucose (cofactor), wild-type GALT catalyzes the conversion of Galactose 1-phosphate Potassium salt to UDP-galactose with a Km value of 0.18 mM (substrate affinity) and a kcat of 25 s⁻¹ (catalytic turnover rate) [1]
2. Metabolic accumulation in GALT-deficient cell models: In fibroblasts isolated from type I galactosemia patients (with GALT gene mutations, e.g., Q188R mutation), incubation with Galactose 1-phosphate Potassium salt (0.5–5 mM) for 24 h led to dose-dependent intracellular accumulation of galactose 1-phosphate. At 2 mM concentration, the accumulation level in mutant fibroblasts was 7.2-fold higher than that in wild-type fibroblasts, while wild-type cells could rapidly metabolize the substrate with minimal accumulation (≤ 10% of mutant cell levels) [1] 3. Impact of GALT mutations on substrate processing: Recombinant GALT with common type I galactosemia-associated mutations (Q188R, K285N, N314D) showed significantly reduced catalytic efficiency toward Galactose 1-phosphate Potassium salt. The Q188R mutant had a Km of 1.2 mM (6.7-fold higher than wild-type, indicating reduced substrate affinity) and a kcat of 1.8 s⁻¹ (13.9-fold lower than wild-type, indicating impaired catalytic turnover), resulting in a 92% decrease in catalytic efficiency (kcat/Km) compared with wild-type GALT [1] |
| Enzyme Assay |
1. Wild-type and mutant GALT catalytic activity assay using Galactose 1-phosphate Potassium salt as substrate: The assay was established in a buffer system (pH 7.4) containing MgCl₂ (cofactor for GALT), UDP-glucose (second substrate), serial concentrations of Galactose 1-phosphate Potassium salt (0.05–5 mM), and purified recombinant wild-type or mutant GALT enzyme. The reaction was initiated by adding GALT and incubated at 37℃ for 10–30 min (timed to ensure linear reaction kinetics). The formation of UDP-galactose (product of the reaction) was quantified by coupling to a secondary reaction system containing UDP-galactose dehydrogenase and NAD⁺, where the production of NADH (detectable by absorbance at 340 nm) was proportional to UDP-galactose levels. The absorbance was measured using a spectrophotometer at 340 nm every 2 min, and the initial reaction rate was calculated to determine Km (substrate affinity) and kcat (catalytic turnover rate) of GALT for the substrate [1]
2. GALT substrate specificity verification assay: The reaction system was modified to replace Galactose 1-phosphate Potassium salt with other phosphorylated sugar analogs (e.g., glucose 1-phosphate, mannose 1-phosphate) at the same concentration (1 mM). The residual catalytic activity of GALT toward these analogs was measured relative to Galactose 1-phosphate Potassium salt, confirming that GALT exhibited > 95% substrate specificity for galactose 1-phosphate (activity toward other analogs < 5% of the substrate) [1] |
| Cell Assay |
1. Intracellular galactose 1-phosphate accumulation assay in GALT-deficient fibroblasts: Fibroblast cells from type I galactosemia patients (Q188R mutation) and normal control individuals were seeded in 6-well plates (2×10⁵ cells/well) and cultured to 80% confluence. The cells were then incubated with serum-free medium containing Galactose 1-phosphate Potassium salt (0.5–5 mM) for 24 h at 37℃ with 5% CO₂. After incubation, cells were harvested, washed with cold PBS, and lysed with perchloric acid to extract intracellular metabolites. The lysate was neutralized with potassium hydroxide, and the concentration of intracellular galactose 1-phosphate was quantified by high-performance liquid chromatography (HPLC) with UV detection (specific wavelength for phosphorylated sugars). The accumulation level was normalized to total cellular protein content to eliminate cell number differences [1]
2. Cell viability assay in GALT-deficient cells treated with Galactose 1-phosphate Potassium salt: The same fibroblast cells (mutant and wild-type) were seeded in 96-well plates (5×10⁴ cells/well) and treated with Galactose 1-phosphate Potassium salt (0–10 mM) for 48 h. A cell viability detection reagent was added and incubated for 2 h at 37℃, and absorbance was measured at the corresponding wavelength. Wild-type fibroblasts maintained > 90% viability even at 10 mM concentration, while mutant fibroblasts showed a dose-dependent decrease in viability (viability reduced to 58% at 10 mM, with an EC50 of 7.5 mM for cytotoxicity) [1] |
| References | |
| Additional Infomation |
1. Galactose-1-phosphate potassium salt is a natural phosphorylated sugar metabolite in the galactose metabolic pathway and is not a therapeutic drug. It is generated from galactose by the action of galactokinase (GALK) in the first step of galactose catabolism [1]. 2. Pathophysiological effects: In type I galactosemia (caused by autosomal recessive GALT deficiency), galactose-1-phosphate potassium salt cannot be effectively converted into UDP-galactose by GALT, leading to its accumulation in various tissues (liver, kidney, brain, erythrocytes). This accumulation is the main cause of the clinical manifestations of type I galactosemia, including liver dysfunction, cognitive impairment and renal tubular damage [1]. 3. Research applications: Galactose-1-phosphate potassium salt is widely used as a substrate in in vitro and cell experiments to assess GALT enzyme activity, screen GALT-deficient samples and study the functional effects of GALT gene mutations associated with type I galactosemia [1].
|
| Molecular Formula |
C6H11O9P.2K
|
|---|---|
| Molecular Weight |
336.32
|
| Exact Mass |
335.941
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| CAS # |
19046-60-7
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| PubChem CID |
87916
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| Appearance |
White to off-white solid powder
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| Hydrogen Bond Donor Count |
4
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| Hydrogen Bond Acceptor Count |
9
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| Rotatable Bond Count |
2
|
| Heavy Atom Count |
18
|
| Complexity |
266
|
| Defined Atom Stereocenter Count |
5
|
| SMILES |
C([C@@H]1[C@@H]([C@@H]([C@H]([C@H](O1)OP(=O)([O-])[O-])O)O)O)O.[K+].[K+]
|
| InChi Key |
KCIDZIIHRGYJAE-YGFYJFDDSA-L
|
| InChi Code |
InChI=1S/C6H13O9P.2K/c7-1-2-3(8)4(9)5(10)6(14-2)15-16(11,12)13;;/h2-10H,1H2,(H2,11,12,13);;/q;2*+1/p-2/t2-,3+,4+,5-,6-;;/m1../s1
|
| Chemical Name |
dipotassium;[(2R,3R,4S,5R,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)oxan-2-yl] phosphate
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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)
|
| Solubility (In Vitro) |
H2O : ~125 mg/mL (~371.67 mM)
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|---|---|
| Solubility (In Vivo) |
Solubility in Formulation 1: 100 mg/mL (297.34 mM) in PBS (add these co-solvents sequentially from left to right, and one by one), clear solution; with sonication.
(Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 2.9734 mL | 14.8668 mL | 29.7336 mL | |
| 5 mM | 0.5947 mL | 2.9734 mL | 5.9467 mL | |
| 10 mM | 0.2973 mL | 1.4867 mL | 2.9734 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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