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| Targets |
pGpG is the linear byproduct of the enzymatic degradation of cyclic diguanosine monophosphate (c-di-GMP). The target is the phosphodiesterase enzymes that break the cyclic dinucleotide. pGpG itself can be used to detect the activity of enzymes, such as oligoribonuclease (Orn), which further degrade pGpG into two GMP molecules. It also serves as a substrate or inhibitor for various bacterial nucleotidases involved in c-di-GMP turnover.
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| ln Vitro |
5'-Phosphoguanylyl-(3',5')-guanosine suppresses the activity of EAL-dependent PDEs (PA2133, PvrR, and pure recombinant RocR) from P. aeruginosa and decreases the rate of c-di-GMP breakdown in cell lysates [2].
pGpG is a standard used in biochemical assays to detect and quantify the activity of enzymes involved in c-di-GMP metabolism. It is used as a substrate to study the activity of oligoribonuclease (Orn), which cleaves pGpG into two GMP molecules. The detection of pGpG consumption or GMP production serves as a direct readout of nuclease activity, allowing for the characterization of these enzymes. |
| ln Vivo |
pGpG is an endogenous metabolite in bacteria. In vivo, it accumulates as a breakdown product of c-di-GMP signaling. The in vivo levels of pGpG are determined by the balance between the activity of phosphodiesterases (which produce it) and nucleotidases (which degrade it). Therefore, measuring pGpG levels in bacterial cell lysates can provide a snapshot of c-di-GMP turnover and the overall state of the signaling pathway.
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| Enzyme Assay |
The standard assay for pGpG is a reversed-phase LC-MS/MS (Liquid Chromatography with Tandem Mass Spectrometry) method. This technique is used to identify and quantify cyclic dinucleotides like c-di-GMP and their linear metabolite pGpG in bacterial samples. The assay involves lysing bacterial cells, extracting the nucleotides, separating them on a C18 column, and detecting them based on their mass-to-charge ratio.
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| Cell Assay |
This compound is a metabolite and not typically used in cell-based assays as a treatment. However, it is used in the lysis and extraction process to measure the in vivo activity of the c-di-GMP pathway. Upon lysis of bacterial cells, intracellular contents are extracted, and pGpG levels are quantified via LC-MS/MS. The ratio of pGpG to c-di-GMP is used as a metric for phosphodiesterase activity within the cell.
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| Animal Protocol |
In vivo experiments using pGpG are not conducted by treating animals with it. Instead, researchers study its role by manipulating the enzymes that produce and degrade it. For instance, in mouse models of bacterial infection, researchers use strains of bacteria with gene knockouts for the enzymes that degrade pGpG, such as oligoribonuclease. By tracking bacterial colonization and host immune response, they can determine the importance of pGpG clearance in bacterial virulence and survival.
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| ADME/Pharmacokinetics |
As an endogenous nucleotide, pGpG is a water-soluble molecule. For in vitro assays, pGpG is typically stored as a dry powder at -20degC and reconstituted in a buffer (e.g., Tris-HCl, HEPES) to a stock concentration. It is stable under standard laboratory conditions. Its concentration in biological samples can be in the low micromolar range and is typically measured in cell lysates or culture supernatants.
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| Toxicity/Toxicokinetics |
As a naturally occurring intermediate molecule in the bacterial c-di-GMP signaling pathway, pGpG is not considered a toxic substance. Its acute toxicity is expected to be low. For research use, standard laboratory safety precautions for handling nucleotides and nucleic acids are sufficient. No specific mammalian toxicity data is required as its primary application is in bacteriology and as an analytical standard.
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| References |
[1]. Nesbitt NM, et, al. Modification of a bi-functional diguanylate cyclase-phosphodiesterase to efficiently produce cyclic diguanylate monophosphate. Biotechnol Rep (Amst). 2015 May 5;7:30-37.
[2]. Cohen D, et, al. Oligoribonuclease is a central feature of cyclic diguanylate signaling in Pseudomonas aeruginosa. Proc Natl Acad Sci U S A. 2015 Sep 8;112(36):11359-64. |
| Additional Infomation |
PGpG is a 5'-phosphate-(3'->5')-dinucleotide. It is the conjugate acid of pGpG(3-).
The cyclic dinucleotide c-di-GMP is a ubiquitous bacterial second messenger that controls various processes including biofilm formation, motility, and virulence. pGpG is the linear intermediate produced when c-di-GMP is hydrolyzed by a phosphodiesterase. This molecule is then further processed by a nuclease, such as oligoribonuclease, to complete the signaling cycle. The study of pGpG is critical to understanding the regulation of c-di-GMP signaling. This compound is sold as a research grade reagent for use in biochemical assays and analytical standards. |
| Molecular Formula |
C20H26N10O15P2
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|---|---|
| Molecular Weight |
708.43
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| Exact Mass |
708.105
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| CAS # |
33008-99-0
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| PubChem CID |
135566597
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| Appearance |
White to off-white solid powder
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| LogP |
-7.8
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| Hydrogen Bond Donor Count |
10
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| Hydrogen Bond Acceptor Count |
19
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| Rotatable Bond Count |
10
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| Heavy Atom Count |
47
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| Complexity |
1420
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| Defined Atom Stereocenter Count |
8
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| SMILES |
C1=NC2=C(N1[C@H]3[C@@H]([C@@H]([C@H](O3)COP(=O)(O)O[C@@H]4[C@H](O[C@H]([C@@H]4O)N5C=NC6=C5N=C(NC6=O)N)COP(=O)(O)O)O)O)N=C(NC2=O)N
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| InChi Key |
ZEHOHLFQOXAZHX-MHARETSRSA-N
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| InChi Code |
InChI=1S/C20H26N10O15P2/c21-19-25-13-7(15(34)27-19)23-3-29(13)17-10(32)9(31)5(43-17)1-42-47(39,40)45-12-6(2-41-46(36,37)38)44-18(11(12)33)30-4-24-8-14(30)26-20(22)28-16(8)35/h3-6,9-12,17-18,31-33H,1-2H2,(H,39,40)(H2,36,37,38)(H3,21,25,27,34)(H3,22,26,28,35)/t5-,6-,9-,10-,11-,12-,17-,18-/m1/s1
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| Chemical Name |
[(2R,3S,4R,5R)-5-(2-amino-6-oxo-1H-purin-9-yl)-3,4-dihydroxyoxolan-2-yl]methyl [(2R,3S,4R,5R)-5-(2-amino-6-oxo-1H-purin-9-yl)-4-hydroxy-2-(phosphonooxymethyl)oxolan-3-yl] hydrogen 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 (e.g. under nitrogen), 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: 50 mg/mL (70.58 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 | 1.4116 mL | 7.0579 mL | 14.1157 mL | |
| 5 mM | 0.2823 mL | 1.4116 mL | 2.8231 mL | |
| 10 mM | 0.1412 mL | 0.7058 mL | 1.4116 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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