Mononucleoside-5’-phosphate

The hydrolysis of polynucleotides by specific nucleases, or by very mild chemical conditions, will produce a mixture of the constituent nucleotides. Furthermore, the conditions of hydrolysis may determine whether a 3 or a 5 nucleotide is obtained. Hydrolysis of RNA with snake venom phosphodiesterase, for example, breaks the linkages at (a) in (11.122) giving a series of mononucleoside 5 phosphates. Alkaline hydrolysis with N NaOH at room tanperature, on the other hand, breaks the linkages at (b), giving initially nucleoside cyclic 2 ,3 phosphates which are further hydrolysed to a mixture of 2 and 3 nucleoside phosphates containing the various bases. 

Nucleoside Acyclic Phosphates. - 2.1.1 Mononucleoside Phosphate Derivatives. A preliminary study on a novel solid phase reagent (Scheme 1) for the capture phosphorylation of nucleosides has been described. The 1 % cross-linked divinylbenzene-polystyrene copolymer, containing cyanoethoxy iV,AT-diisop-ropyl phosphine was used for the selective phosphorylation of uridine to 5 -uridine monophosphate (UMP) in 67% yield. ... 

The mononucleotides (mononucleoside phosphates) are obtainable by breaking down the polynucleotides or by phosphorylation of pre-formed nucleosides. Their main biochemical role is to function as sources of the nucleoside pyro- and tri-phosphates (below). 

Example 40 mononucleoside phosphotriester derivatives with 5-acyl-2-thioethyl bioreversible phosphate-protecting groups have been synthesized by Imbach and his associates [74a,b] in their research on intracellular delivery of 3 -azido-3 -dideoxythymidine-5 -mono-phosphate. 

Lambda-exonuclease treatment. This procedure is used by Alderon Biosciences [28]. Exonucleases are enzymes that hydrolyse the terminal phosphodiester bonds of DNA strands obtaining mononucleosides monophosphate or biphosphate. In this case a 5 -exo-nuclease that hydrolyses the 5 -end phosphodiester is used. In this way, the strands are destroyed. To achieve the total hydrolysis of the sister strand, the target strand must be protected, for example, by a label at its 5 -end. Therefore, the PCR amplification is carried out using one of the primers biotinylated at the 5 -end and the other one modified at its 5 -end with a phosphate group. The extension of these primers give rise to the 5 -biotinylated target DNA strand and a 5 -phosphate sister DNA strand, which is hydrolysed by 5 -exonuclease. 

Gorenstein Unfortunately I do not know of any chemical shift anisotropy theoretical studies on ATP (or any phosphates in fact). Some recent advances in solid state, oriented crystal, P NNR of various nucleic acids have appeared in the literature. Large C.S.A. s are found for mononucLeoside monophosphates. Your suggestion for the line broadening in ATP is dertainly reasonable but possibly could also be explained by chemical exchange broadening ( ). 

Phosphate Activator. MST was shown to be an effective catalyst for the phosphorylation of a 5 -protected mononucleoside. MST was stirred with /t-chlorophenyl phosphorodichloridate and triethylamine and the resulting agent was used directly for the phosphorylation of 5 -protected nucleosides in 70-85% yields. 

Narang and Michniewicz (87) reported successful application of Avicel-cellulose plates for the rapid and efficient fractionation of complex chemically polymerized thymidine 5 -phosphate reaction mixtures, in situ quantitative analysis of the oligonucleotides on TLC plate by reflectant spectrophotometry, in situ identiflcation of the common mononucleosides and mononucleotides by double-scanning technique and fractionation of oligonucleotides on thick-layer Avicel-cellulose plates on a preparative scale. 

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