Synthetases diphosphate synthetase

Whereas imidazolides of nucleotides react only in organic solvents with phosphates or pyrophosphates to give the corresponding anhydride derivatives in high yield, ATP can also be formed enzymatically in aqueous solution from AMP-Im with inorganic pyrophosphate in the presence of valyl-f-RNA synthetase.[66] A variant of this method is the one-pot reaction of a nucleoside with phosphoryltristriazole and tributylammonium pyrophosphate. 671 An a-methylphosphonyl-/ ,y-diphosphate of a thymidine derivative has been synthesized in a similar way t681... 

The above reactions are shown schematically in Fig. 5.1. The overall reaction is reversible but is, however, shifted to the right, since the enzyme pyrophospho-rylase hydrolyses the diphosphate formed, thus removing it from the equilibrium. Each of the 20 proteinogenic amino acids has at least one specific synthetase. In steps 1 and 2, the amino acid exists in two different forms—in step 1 as a mixed anhydride ... 

The hypE proteins are 302-376 residues long and appear to consist of three domains. Domain 1 shows sequence identity to a domain from phosphoribosyl-aminoimida-zole synthetase which is involved in the fifth step in de novo purine biosynthesis and to a domain in thiamine phosphate kinase which is involved in the synthesis of the cofactor thiamine diphosphate (TDP). TDP is required by enzymes which cleave the bond adjacent to carbonyl groups, e.g. phosphoketolase, transketolase or pyruvate decarboxylase. Domain 2 also shows identity to a domain found in thiamine phosphate kinase. Domain 3 appears to be unique to the HypF proteins. 

The subsequent cleavage of the thio-ester succinylCoA into succinate and coenzyme A by succinic acid-CoA ligase (succinyl CoA synthetase, succinic thiokinase) is strongly exergonic and is used to synthesize a phosphoric acid anhydride bond ( substrate level phosphorylation , see p. 124). However, it is not ATP that is produced here as is otherwise usually the case, but instead guanosine triphosphate (CTP). However, GTP can be converted into ATP by a nucleoside diphosphate kinase (not shown). 

This enzyme [EC 6.2.1.15], also known as arachido-nateiCoA ligase, catalyzes the reaction of arachidonate with ATP and coenzyme A to generate arachidonyl-CoA, AMP, and pyrophosphate (or, diphosphate). The enzyme can also use 8,11,14-icosatrienoate as a substrate, but not the other long-chain fatty acids. It should be noted that this enzyme is not identical to long-chain acyl-CoA synthetase [EC 6.2.1.3]. 

This enzyme [EC 6.1.1.12], also known as aspar-tate tRNA ligase, catalyzes the reaction of aspartate with ATP and tRNA P to generate aspartyl-tRNA P, AMP, and diphosphate (or, pyrophosphate). See also Amino-acyl-RNA Synthetases... 

GMP synthetase [EC 6.3.4.1], also known as xanthosine-5 -phosphate ammonia ligase, catalyzes the reaction of ATP with xanthosine 5 -phosphate and ammonia to produce GMP, AMP, and pyrophosphate (or, diphosphate). GMP synthetase (glutamine-utilizing) [EC 6.3.5.2] catalyzes the reaction of ATP with xanthosine 5 -phosphate, L-glutamine, and water to produce GMP, AMP, L-glutamate, and pyrophosphate. 

AMP, and pyrophosphate (or, diphosphate). Propenoate can also act as the substrate. This enzyme is not identical with acetyl-CoA synthetase or with butyryl-CoA synthetase. 

This enzyme [EC 2.7.6.1], also known as phosphoribosyl pyrophosphate synthetase, catalyzes the reaction of ATP with D-ribose 5-phosphate to produce AMP and 5-phospho-a-D-ribose 1-diphosphate. dATP can also function as a substrate. 

This dependence on light levels and temperature is believed to be due to the mechanism of production of isoprene in the plant, which involves the enzyme isoprene synthetase and dimethylallyl diphosphate (DMAPP) as a precursor to isoprene (e.g., see Silver and Fall, 1995 and Monson et al., 1995). Either the enzyme, the formation of DMAPP, or both may be light sensitive (Wildermuth and Fall, 1996). The temperature effect has been attributed to effects on the enzyme, increasing its activity initially and then leading to irreversible denaturation (and/or possibly membrane damage) (Fall and Wildermuth, 1998). 

Kavanaugh-Black, A. Connolly, D.M. Chugani, S.A. Chakrabarty, M. Characterization of nucleoside-diphosphate kinase from Pseudomonas aeruginosa complex formation with succinyl-CoA synthetase. Proc. Natl. Acad. Sci. USA, 91, 5883-5887 (1994)... 

Kowluru, A. Tannous, M. Chen, H.Q. Localization and characterization of the mitochondrial isoform of the nucleoside diphosphate kinase in the pancreatic p cell evidence for its complexation with mitochondrial succinyl-CoA synthetase. Arch. Biochem. Biophys., 398, 160-169 (2002)... 

The formation of ATP (or GTP) at the expense of the energy released by the oxidative decarboxylation of a-ketoglutarate is a substrate-level phosphorylation, like the synthesis of ATP in the glycolytic reactions catalyzed by glyceraldehyde 3-phosphate dehydrogenase and pyruvate kinase (see Fig. 14-2). The GTP formed by succinyl-CoA synthetase can donate its terminal phosphoryl group to ADP to form ATP, in a reversible reaction catalyzed by nucleoside diphosphate kinase (p. 505) ... 

Thus the net result of the activity of either isozyme of succinyl-CoA synthetase is the conservation of energy as ATP. There is no change in free energy for the nucleoside diphosphate kinase reaction ATP and GTP are energetically equivalent. 

Stoichiometric quantities of uridine diphosphate glucose were used, in the presence of a transfer enzyme, sucrose synthetase, in the soluble state (extraction given). Coupling with modified D-fructose gave sucroses modified on the D-fructosyl group, on the 1 -3-mmol scale. Thus were prepared l -deoxy-l -fluoro- (59%),98 4 -deoxy-4 -fluoro- (16%), and l -azido-l -deoxy-sucrose (15%).71 6-Deoxy-6-fluoro-D-glucose was isomerized to 6-deoxy-6-fluoro-D-fructose with isomerase, and gave 6 -deoxy-6 -fluoro-sucrose.71... 

Didanosine is a synthetic purine nucleoside analog that inhibits the activity of reverse transcriptase in HIV-1, HIV-2, other retroviruses and zidovudine-resistant strains. A nucleobase carrier helps transport it into the cell where it needs to be phosphorylated by 5 -nucleoiidase and inosine 5 -monophosphate phosphotransferase to didanosine S -monophosphate. Adenylosuccinate synthetase and adenylosuccinate lyase then convert didanosine 5 -monophosphate to dideoxyadenosine S -monophosphate, followed by its conversion to diphosphate by adenylate kinase and phosphoribosyl pyrophosphate synthetase, which is then phosphorylated by creatine kinase and phosphoribosyl pyrophosphate synthetase to dideoxyadenosine S -triphosphate, the active reverse transcriptase inhibitor. Dideoxyadenosine triphosphate inhibits the activity of HIV reverse transcriptase by competing with the natural substrate, deoxyadenosine triphosphate, and its incorporation into viral DNA causes termination of viral DNA chain elongation. It is 10-100-fold less potent than zidovudine in its antiviral activity, but is more active than zidovudine in nondividing and quiescent cells. At clinically relevant doses, it is not toxic to hematopoietic precursor cells or lymphocytes, and the resistance to the drug results from site-directed mutagenesis at codons 65 and 74 of viral reverse transcriptase. 

Animals are unable to synthesize folic acid (6.62) and must consume adequate quantities in their diets. Plants and bacteria, however, are able to make folic acid. The first step of this synthesis is catalyzed by dihydropteroate synthetase and reacts dihydroptero-ate diphosphate (6.69) and para-aminobenzoic acid (PABA, 6.70) (Figure 6.25). Because this pathway is not found in humans, inhibition of the reaction is a method to ultimately stop TMP synthesis in an invading bacterium while not impacting the infected host. The sulfonamides, often called sulfa drugs, are a class of antibiotic that exploits the folic acid pathway and inhibits dihydropteroate synthetase. Sulfa drugs bind in the same fashion as PABA and act as competitive inhibitors. The active form of the first sulfa drug is sulfanilamide (6.71). Sulfamethoxazole (6.72) is a sulfa drug that is widely prescribed today.26... 

Some of the reactions of PO3- parallel enzymatic reactions promoted by adenosine triphosphate (ATP). Pyruvate kinase catalyzes the equilibration of ATP and pyruvate with adenosine diphosphate (ADP) and phosphoenol pyruvate (11,12). In a formal sense, this reaction resembles the preparations of enol phosphate (eqs. 6 and 7). Cytidine triphosphate synthetase catalyzes the reaction of uridine triphosphate with ammonia to yield cytidine triphosphate (13). In a formal sense, this reaction resembles the replacement of the ester carbonyl group of ethyl acetate by the nitrogen of aniline (eq. 8). 

Fig. 15-15 The de novo pyrimidine biosynthetic pathway. CAP, carbamoyl phosphate CA-asp, /V-carbamoyl-L-aspartate DHO, L-dihydroorotate Oro, orotate OMP, orotidine 5 -monophosphate. Enzymes (1) carbamoyl phosphate synthetase II (2) aspartate transcarbamoylase (3) dihydroorotase, (4) dihydroorotate dehydrogenase (5) orotate phosphoribosyltransferase (6) OMP decarboxylase (7) nucleoside monophosphate kinase (8) nucleoside diphosphate kinase (9) CTP synthetase.

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