Nucleoside 5 -diphosphate

Synthetic oligonucleotides may be used as "primers and be elongated stepwise with the aid of polynucleotide phosphorylase (PNPase) and nucleoside diphosphates. 

FIGURE 23.6 The PEP carboxykinase reaction. GTP formed in this reaction can be converted to ATP by nucleoside diphosphate kinase, althongh liver cells in some species may not contain this enzyme. 

The NDP-dependent potassium (KndP) channel is activated by nucleoside diphosphates (NDPs) in the presence of Mg2+ and is relatively insensitive to inhibition by ATP. Kndp channels are present in vascular smooth muscle. 

By means of the enzyme nucleoside diphosphate kinase, UTP, GTP, and CTP can be synthesized from their diphosphates, eg,... 

All of these triphosphates take part in phosphorylations in the cell. Similarly, specific nucleoside monophosphate kinases catalyze the formation of nucleoside diphosphates from the corresponding monophosphates. 

Other nucleoside diphosphate sugar compounds are known, eg, UDPGal. In addition, the same sugar may be linked to different nucleotides. For example, glucose may be linked to uridine (as shown above) as well as to guanosine, thymidine, adenosine, or cy-tidine nucleotides. 

A primed numeral locates the position of the phosphate on the sugars of mononucleotides (eg, 3 -GMP, 5 -dCMP). Additional phosphoryi groups linked to the first by acid anhydride bonds form nucleoside diphosphates and triphosphates. 

While mammahan cells reutilize few free pyrimidines, salvage reactions convert the ribonucleosides uridine and cytidine and the deoxyribonucleosides thymidine and deoxycytidine to their respective nucleotides. ATP-dependent phosphoryltransferases (kinases) catalyze the phosphorylation of the nucleoside diphosphates 2 "-de-oxycytidine, 2 -deoxyguanosine, and 2 -deoxyadenosine to their corresponding nucleoside triphosphates. In addition, orotate phosphoribosyltransferase (reaction 5, Figure 34-7), an enzyme of pyrimidine nucleotide synthesis, salvages orotic acid by converting it to orotidine monophosphate (OMP). 

C. Nucleoside Diphosphate Sugars.—A polyprenol phosphate containing eleven isoprene units is involved in the biosynthesis of various bacterial cell-wall components.As mentioned in last year s Report, another isoprenoid phosphate, dolichol monophosphate (40), is an intermediate in sugar... 

NCF Neutrophil chemotactic factor NDGA Nordihydroguaretic acid NDP Nucleoside diphosphate Neca 5 -(N-ethyl carboxamido)-adenosine NED Nedocromil sodium NEP Neutral endopeptidase (EC 3.4.24.11)... 

Nucleoside triphosphates are prepared analogously to the diphosphates by the methods a, b, c, and d, representing conversion of a nucleoside phosphoric azolide with inorganic pyrophosphate (Method a), of a nucleoside diphosphoric azolide with inorganic phosphate (Method b), of a phosphoric bisazolide with a nucleoside diphosphate (Method c), and of a diphosphoric bisazolide (bisazolide of pyrophosphoric acid) with a nucleoside phosphate (Method d). 

Another route to such nucleoside diphosphates starts from imidazolides of phosphoric acid monoester and nucleoside phosphates ... 

Handschuh and Orgel (1973) studied the mineral struvite. It can be precipitated from ocean water in the presence of phosphate if the concentration of NH ions in the water is greater than 0.01 M. If struvite is heated with urea, magnesium pyrophosphate is obtained in a yield of about 20% after 10 days at 338 K if nucleosides are added to the reaction mixture described above, nucleoside diphosphates such as uridine-5 -diphosphate and diuridine-5 -diphosphate are formed in good yields. 

Deoxynucleotides for DNA synthesis are made at the nucleoside diphosphate level and then have to be phosphorylated up to the triphosphate using a kinase and ATP. The reducing equivalents for the reaction come from a small protein, thioredoxin, that contains an active site with two cysteine residues. Upon reduction of the ribose to the 2 -deoxyri-bose, the thioredoxin is oxidized to the disulfide. The thioredoxin(SS) made during the reaction is recycled by reduction with NADPH by the enzyme thioredoxin reductase. 

UDP and UTP are selective agonists of certain of the P2Y receptors. It is not yet clear what factors control the release of uridine nucleotides into the extracellular space. UTP can be formed from UDP in the extracellular space by the action of the enzyme nucleoside diphosphokinase, which catalyzes the transfer of the -phosphate of nucleoside triphosphates to nucleoside diphosphates, e.g., ATP + UDP —> ADP + UTP. 

The Fe-protein has the protein fold and nucleotide-binding domain of the G-protein family of nucleotide-dependent switch proteins, which are able to change their conformation dependent on whether a nucleoside diphosphate (such as GDP or ADP) is bound instead of the corresponding triphosphate (GTP or ATP). However, nucleotide analogues, which induce the conformational switch of the Fe-protein, do not allow substrate reduction by the MoFe-protein, nor does reduction of the MoFe-protein by other electron-transfer reagents (whether small proteins or redox dyes) drive substrate reduction. Only the Fe-protein can reduce the MoFe-protein to a level that allows it to reduce substrates such as... 

DNA and RNA synthesis De novo formation of purine and pyrimidine nucleotide Nucleoside diphosphate reductase Thymidylate synthase Polymerase reactions Chapter 20... 

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