| Size | Price | |
|---|---|---|
| Other Sizes |
Purity: ≥98%
| Targets |
γ-glutamyl cycle (possible involvement) [1]
|
|---|---|
| ln Vitro |
The uptake of glycyl-L-leucine by monkey small intestinal strips involves two saturable transport processes. One process has a maximum velocity (Vmax) of 1 μmol min⁻¹ g⁻¹ wet weight of tissue and an affinity constant (kt) of 5 mmol/l. The other process has a Vmax of 3.9 μmol min⁻¹ g⁻¹ wet weight of tissue and a kt of 33 mmol/l. [1]
The uptake of glycyl-L-[¹⁴C]leucine is inhibited by a wide variety of amino acids. At a concentration of 50 mmol/l, the percentage inhibition by various L-amino acids is as follows: Phenylalanine (62.2%), Alanine (59.0%), Methionine (58.4%), Leucine (56.1%), Cysteine (53.2%), Lysine (49.4%), Threonine (47.1%), Valine (43.4%), Histidine (42.1%), Glutamine (42.0%), Tryptophan (31.3%), Proline (30.2%), Arginine (29.9%), Serine (28.2%), Glycine (27.0%), Hydroxyproline (26.3%), Asparagine (24.2%), Glutamic acid (6.6%), Aspartic acid (4.8%). [1] The inhibition of glycyl-L-leucine uptake by the amino acid leucine was shown to be competitive. This was demonstrated by varying the dipeptide concentration (5-50 mmol/l) in the presence of 50 mmol/l leucine. [1] The effect of various amino acids (40 mmol/l) on the hydrolysis of glycyl-L-leucine (8 mmol/l) by cytosol and particulate (brush-border) fractions of monkey intestinal mucosa was studied. Cysteine completely inhibited both the cytosol and particulate hydrolase activities. Histidine strongly inhibited both (95.4% for cytosol, 73.4% for particulate). Several amino acids (e.g., Methionine, Leucine, Norleucine, Tryptophan, Phenylalanine, Isoleucine) showed greater inhibition of the cytosol enzyme (ranging from 59.6% to 73.2%) compared to the particulate enzyme (ranging from 22.3% to 47.2%). Glycine, proline, alanine, and glutamic acid did not inhibit either enzyme activity. [1] There is no correlation between the effects of amino acids on glycyl-L-leucine uptake and on its hydrolysis by either the particulate or cytosol fraction. For example, alanine inhibits uptake (59.0%) but does not inhibit either hydrolase; aspartic acid inhibits both hydrolases (44.8% for cytosol, 38.3% for particulate) but does not inhibit uptake (4.8%). [1] |
| Enzyme Assay |
Glycyl-L-leucine hydrolase activity was assayed using a modified chromatographic method. The standard assay mixture contained 2.5 μmol of glycyl-L-leucine, 5 μmol of Tris/HCl buffer (pH 7.8), and the enzyme in a total volume of 0.05 ml. Incubation was carried out at 37°C for 5 minutes. The reaction was stopped by placing the mixture in a boiling-water bath for 1 minute. Aliquots of the assay mixtures were spotted on Whatman no. 1 paper, and descending chromatography was developed with propan-2-ol/water (4:1, v/v). After air-drying, the paper was sprayed with 0.5% ninhydrin in acetone/water (19:1, v/v). The spots containing leucine were cut out and eluted with 4 ml of ethanol/water (3:1, v/v). 0.04 ml of copper sulphate solution (3 g/100 ml) in water was added, and the color intensity was read at 510 nm. This method quantified leucine in the range of 0.01-0.125 μmol. [1]
To specifically assay brush-border peptide hydrolase activity, the assay mixture contained 0.1 mmol/l p-hydroxymercuribenzoate, which was shown to inhibit cytosol peptide hydrolases by 92-100% without affecting brush-border enzymes. [1] |
| Cell Assay |
Uptake experiments were performed in vitro using everted strips of monkey small intestine. After the monkeys were sacrificed, the small intestine was removed, washed with ice-cold oxygenated Krebs-Ringer bicarbonate buffer (pH 7.4) containing 5% glucose, and everted. Rectangular strips (whole wall, weighing 30-40 mg) from a specific region (35-40% of the total length from the pyloric end) were used. Each strip was incubated with 0.5 ml of the dipeptide solution in Krebs-Ringer bicarbonate buffer (pH 7.4) with 5% glucose. D-Mannitol was used to equalize osmolality. Incubations were carried out at 37°C for 2 minutes. After incubation, the strip was blotted, weighed, and digested in 0.5 ml of 2 mol/l NaOH at 75°C for 2 hours. The digested tissue was mixed with a scintillation mixture (containing naphthalene, 2,5-diphenyloxazole, and Cab-o-Sil in dioxan), and the radioactivity was counted. The extracellular space was determined separately using [³H]inulin and was found to be 5% of the wet weight of the tissue for a 2-minute incubation. The average total water content was 78 ± 3% of the wet weight. [1]
|
| References | |
| Additional Infomation |
Glycyl-Leu is a dipeptide composed of glycine and L-leucine linked by a peptide bond; it is a metabolite. Its function is related to glycine and L-leucine, and it is a zwitterion tautomer of glycyl-L-leucine. Glycyl-L-leucine has been reported to exist in fruit flies, humans, and several other organisms with relevant data.
In the mammalian intestine, the transport of peptides may be physiologically and nutritionally more important than the transport of amino acids. However, studies suggest these two transport systems may interact, with dipeptides and amino acids possibly competing during transport. [1] The inhibition of dipeptide uptake by free amino acids has been hypothesized to occur via retardation of dipeptide hydrolysis, leading to increased intracellular peptide concentration and a secondary decrease in the entry gradient. However, the lack of correlation between uptake inhibition and hydrolase inhibition in this study suggests a direct interaction of amino acids with the dipeptide transport system. [1] The study also discusses the potential role of the γ-glutamyl cycle in the transport of amino acids and peptides, as γ-glutamyl transpeptidase can utilize both amino acids and dipeptides as acceptors. The observed mutual interaction and lack of amino acid group specificity in this study are seen as consistent with this hypothesis. [1] |
| Molecular Formula |
C8H16N2O3
|
|---|---|
| Molecular Weight |
188.22
|
| Exact Mass |
188.116
|
| CAS # |
869-19-2
|
| Related CAS # |
869-19-2;
|
| PubChem CID |
92843
|
| Appearance |
White to off-white solid powder
|
| Density |
1.1±0.1 g/cm3
|
| Boiling Point |
410.4±30.0 °C at 760 mmHg
|
| Melting Point |
233-235 °C(lit.)
|
| Flash Point |
202.0±24.6 °C
|
| Vapour Pressure |
0.0±2.1 mmHg at 25°C
|
| Index of Refraction |
1.489
|
| LogP |
-0.09
|
| Hydrogen Bond Donor Count |
3
|
| Hydrogen Bond Acceptor Count |
4
|
| Rotatable Bond Count |
5
|
| Heavy Atom Count |
13
|
| Complexity |
192
|
| Defined Atom Stereocenter Count |
1
|
| SMILES |
[C@@H](C(=O)O)(CC(C)C)NC(=O)CN
|
| InChi Key |
DKEXFJVMVGETOO-LURJTMIESA-N
|
| InChi Code |
InChI=1S/C8H16N2O3/c1-5(2)3-6(8(12)13)10-7(11)4-9/h5-6H,3-4,9H2,1-2H3,(H,10,11)(H,12,13)/t6-/m0/s1
|
| Chemical Name |
glycyl-L-leucine
|
| Synonyms |
Glycylleucine N-Glycyl-L-leucine NSC 83257 NSC-83257 NSC83257
|
| HS Tariff Code |
2934.99.9001
|
| 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)
|
| Solubility (In Vitro) |
H2O : ~25 mg/mL (~132.82 mM)
|
|---|---|
| Solubility (In Vivo) |
Solubility in Formulation 1: 50 mg/mL (265.65 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 | 5.3129 mL | 26.5647 mL | 53.1293 mL | |
| 5 mM | 1.0626 mL | 5.3129 mL | 10.6259 mL | |
| 10 mM | 0.5313 mL | 2.6565 mL | 5.3129 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.
| NCT Number | Recruitment | interventions | Conditions | Sponsor/Collaborators | Start Date | Phases |
| NCT03496961 | UNKNOWN STATUS | Behavioral: Yan Nian Jiu Zhuan Fa | Chronic Fatigue Syndromes | Shanghai University of Traditional Chinese Medicine | 2018-12-01 | Not Applicable |
Products are for research use only; We do not sell to patients
Copyright 2020 InvivoChem LLC | All Rights Reserved
COA