N⁶-Propargyl-ATP (N⁶pATP)

For general laboratory use.

Shipping: shipped on gel packs

Storage Conditions: store at -20 °C
Short term exposure (up to 1 week cumulative) to ambient temperature possible.

Shelf Life: 12 months after date of delivery

Molecular Formula: C₁₃H₁₈N₅O₁₃P₃ (free acid)

Molecular Weight: 545.23 g/mol (free acid)

Exact Mass: 545.01 g/mol (free acid)

Purity: ≥ 95 % (HPLC)

Form: solid

Color: white to off-white

Solubility: 10 mM Tris-HCl pH 7.5

Spectroscopic Properties: λ_max 262 nm, ε 18.0 L mmol^-1 cm^-1 (Tris-HCl pH 7.5)

Applications:
in vitro AMPylation of proteins^[1,2]
in vitro polyadenylation of RNA^[3]
The resulting alkyne-functionalized protein^[1,2] or RNA^[3] can subsequently be processed via Cu(I)-catalyzed (azide-alkyne) click chemistry that offers the choice

• to introduce a Biotin group for subsequent purification tasks (via Azides of Biotin)
• to introduce fluorescent group for subsequent microscopic imaging (via Azides of fluorescent dyes)
• to crosslink the RNA to azide-functionalized biomolecules e.g.proteins

Presolski et al.^[4] and Hong et al.^[5] provide a general protocol for Cu(I)-catalyzed click chemistry reactions that may be used as a starting point for the set up and optimization of individual assays.
Agonistic ligand, mainly for nucleoside receptor A₁
Nucleoside-triphosphates can be converted by different membrane-bound phosphatases into nucleosides acting as nucleoside receptor ligands. In some cases nucleoside phosphates act also directly on nucleoside receptors.

Please note: This compound contains a phosphoramide linkage which is hydrolyzed at pH <7.0.
For preparation of a 10 mM solution use 100 mM buffer (for example: bicarbonate, borate, phosphate and Tris) to prevent degradation at acidic pH.

Related products:

• Copper (II)-Sulphate (CuSO₄), #CLK-MI004
• Tris(3-hydroxypropyltriazolylmethyl)amine (THPTA), #CLK-1010
• Sodium Ascorbate (Na-Ascorbate), #CLK-MI005

Selected References:
[1] Grammel et al. (2011) A Chemical Reporter for Protein AMPylation. J. Am. Chem. Soc. 133:17103.
[2] Broncel et al. (2012) A New Chemical Handle for Protein AMPylation at the Host-Pathogen Interface. ChemBioChem 13:183.
[3] Grammel et al. (2012) Chemical Reporter for Monitoring RNA Synthesis and Poly (A) Tail Dynamics. ChemBioChem 13:1112.
[4] Presolski et al. (2011) Copper-Catalyzed Azide-Alkyne Click Chemistry for Bioconjugation. Current Protocols in Chemical Biology 3:153.
[5] Hong et al. (2011) Analysis and Optimization of Copper-Catalyzed Azide-Alkyne Cycloaddition for Bioconjugation. Angew. Chem. Int. Ed. 48:9879.
Sirci et al. (2012) Ligand-, structure- and pharmacophore-based molecular fingerprints: a case study on adenosine A1, A2A, A2B, and A3 receptor antagonists. J. Comput. Aided Mol. Des. 26:1247.
Volonte et al. (2009) Membrane components and purinergic signalling: the purinome, a complex interplay among ligands, degrading enzymes, receptors and transporters. FEBS J. 276:318.
Yegutkin (2008) Nucleotide and nucleoside converting enzymes: Important modulators of purinergic signalling cascade. Biochim. Biophys. Acta 1783:673.
Joshi et al. (2005) Purine derivatives as ligands for A3 adenosine receptors. Current Topics in Medicinal Chemistry 5:1275.
Volpini et al. (2002) N6-Alkyl-2-alkynyl derivatives of adenosine as potent and selective agonists of the human adenosine A3 receptor and starting point for searching A2B ligands. J. Med. Chem. 45 (15):3271.
Hess (2001) Recent advantages in adenosine receptor antagonist research. Expert Opin. Ther. Patents 11 (10):1533.
Jacobson (2001) Probing adenosine and P2 receptors: design of novel purines and nonpurines as selective ligands. Drug Development Res. 52:178.
Jacobson et al. (2001) Ribose modified nucleosides and nucleotides as ligands for purine receptors. Nucleosides, Nucleotides & Nucleic Acids 20 (4):333.
Van Galen et al. (1994) A binding site model and structure-activity relationships for rat A3 adenosine receptor. Molecular Pharmacology 45:1101.