Adenosine-5-phosphosulfate

ATP interacts with sulfate ion (SO ) to form adenosine 5-phosphosulfate (APS) in a reaction mediated by the enzyme ATP sulfate adenylyl transferase. 

L-cysteine. The process is completed by glucosylation and addition of S03 at the nitrogen (Equation 10) by reaction with 3 -phospho-adenosine-5 -phosphosulfate. 

Lyric RM, Suzuki I. 1970. Enzymes involved in the metabolism of thiosulfate by Thiobacillus thioparus. II. Properties of adenosine 5 -phosphosulfate reductase. Can J Biochem 48 344-54. 

Peck HD. 1960. Adenosine 5 -phosphosulfate as an intermediate in the oxidation of thiosulfate by Thiobacillus thioparus. Proc Nat Acad Sci USA 46 1053-7. 

Peck HD, Deacon TE, Davidson JT. 1965. Studies on adenosine 5 -phosphosulfate reductase from Desulfovibrio desulfuricans and Thiobacillus thioparus. Biochim Biophys Acta 96 429 7. 

ADP NADH Disappearance Pyruvate Kinase, Lactate Dehydrogenase, and 3 -Nucleo-tidase Adenosine-5 -phosphosulfate Kinase ... 

Oxidation of sulfite to sulfate within cells occurs by a pathway through adenosine 5 -pHosphosulfate (APS, adenylyl sulfate). Oxidation via APS (Eq. 18-22) provides a means of substrate-level phosphorylation,... 

Figure 2. Structure of the adenosine-5 -phosphosulfate (APS) molecule (Reproduced with permission from Ref. 9. Copyright 1986 U. Fischer).

Kinases and sulfotransferases utilize similar substrates and catalyze similar reactions. Both transfer anionic groups (Scheme 14.9). Both enzyme classes are capable of binding adenosine-based substrates. Sulfotransferases bind 3 -phospho-adenosine-5 -phosphosulfate (PAPS) (35) as a sulfate donor and kinases bind adenosine-5 -triphosphate (ATP) (36) as a phosphoryl donor. 

ATP-sulfurylase (ATP sulfate adenyltransferase) catalyzes the synthesis of adenosine 5 -phosphosulfate, using sulfate and ATP as substrates. This is the... 

Adenosine 5 -phosphosulfate was separated from ATP and other adenine nucleotides by chromatography on a Synchropak AX-100 column (4.1 mm x 250 mm). The mobile phase contained 0.1 M sodium phosphate buffer (pH 7.3) and 0.8 M NaHC03. The effluent profile was obtained by monitoring at 254 nm. 

Figure 9.141 Time course of formation of adenosine 5 -phosphosulfate (APS) by activity in rat liver. The reaction mixture contained, in a final volume of 350 /xL, 30 ju.mol of Tris-HCl (pH 8.0), 0.9 /xmol of ATP, 3 /unol of magnesium sulfate, 6 /xmol of sodium fluoride, and 50 fiL of inorganic pyrophosphatase (2.5 U). A50/xL supernatant sample (19 mg of protein) from rat liver was added to start the reaction. Then samples were removed at intervals, and the reaction was terminated and analyzed. Chromatograms were obtained after incubation for (A) 0 minutes, (B) 15 minutes, and (C) 45 minutes. Arrow indicates elution time for the reaction product APS. (From Mina and Rossomando, 1988.)...

Two major pathways are known for the reduction of sulfate. One is the assimilatory pathway, which reduces sulfate to the extent necessary for satisfying the nutritional requirements of the organism. In this pathway, which has been extensively studied in yeast by Robbins and Lip-mann (S68) and Bandurski and his colleagues 369, 370), sulfate is first activated in the presence of ATP by the enzyme ATP-sulfurylase to form adenosine 5 -phosphosulfate (APS). Then in a second reaction, APS is phosphorylated in the 3 position by ATP to form 3 -phosphoadenosine 5 -phosphosulfate (PAPS)... 

As pointed out in the preceding section, sulfate assimilation in yeast has been shown to involve the activation of sulfate by ATP successively to adenosine 5 -phosphosulfate and once again to 3 -phosphoadenosine 5 -phosphosulfate. The latter is then reduced in the presence of NADPH to sulfite and 3, 5 -diphosphoadenosine (37 ). Enzymes catalyzing the... 

A.7.1 Esterification of Acids using Carbodiimides. The formation of anhydrides from carboxylic acids, thiocarboxylic acids, sulfonic acids and phosphorous acids are discussed in Section 2.4.S.2. In this section only special cases of anhydride formation are described. Mixed anhydrides of amino acids and adenylic acid are produced from the corresponding acids using DCC as the condensation agent. ° Mixed anhydrides not containing amino acids, such as butyryl adenate, adenosine 5 -phosphosulfate and p-nitrophenyl-thymidine-5-phosphate are also obtained. 

Nucleoside Pyrophosphates. - 2.2.1 Nucleoside Diphosphate Analogues. The enzymatic regeneration of 3 -phosphoadenosine-5 -phosphosulfate (76) catalysed by the rat liver sulfotransferase IV enzyme has been reported to provide useful quantities of the cofactor for enzyme kinetic experiments. The first thio-nucleotide analogues of adenosine 5 -phosphosulfate (77a) and of 3 -phosphoadenosine 5 -phosphosulfate (77b) have been synthesised in enantio-merically pure forms. Using these novel analogues, the sulfuryl transfer reaction to adenosine 5 -triphosphate was shown to proceed with inversion of configuration at the a-phosphorus. ... 

Pyrosequencing is a method to determine the nucleic acid sequence of short segments without the use of electrophoresis. A sequencing primer is hybridized to a single-stranded template that is usually generated by PCR. Four enzymes, a DNA polymerase, ATP sulfurylase, luciferase and apyrase, and two substrates— adenosine 5 phosphosulfate and luciferin— are included in the reaction mixture (Figure 37-17). One of the four dNTPs is added to the reaction (dATPaS is substituted for dATP because it is incorporated by the polymerase but is not a luciferase substrate). If the base is complementary to the template strand, DNA polymerase catalyzes its incorporation. Each incorporation event is accompanied by release of a pyrophosphate (PPi) so that the quantity of PPi produced is equimolar to the... 

Figure 37-17 Schematic of pyrosequencing. individual dTNPs are added one by one to the single-stranded template, a primer, and a polymerase. Pyrophosphate is generated if the dNTP is complementary to the next base on the template (top). Any pyrophosphate produced reacts with adenosine-5 -phosphosulfate (APS) to produce ATP, which in turn generates light in the presence of Iuciferase (middle). The sequence can be determined from the order of dTNP addition and the intensity of light produced.

NMR, nuclear magnetic resonance mV, millivolt pCMB, p-chloromercuribenzoate pCMS, p-chloromercuriphenyl sulfonate PMS, phenazine methosulfate EDTA, ethylenediaminetetraacetate SDS, sodium dodecyl sulfate TTFA, 2-thenoyltriflu-oroacetone ETP, submitochondrial (electron transfer) particles MVH, reduced methyl viologen DEAE-cellulose, diethylaminoethyl cellulose succ, succinate APS, adenosine 5 -phosphosulfate PAPS, 3 -phosphoadenosine 5 -phosphosulfate. Other abbreviations are standard [see JBC 244, 2 (1969)]. 

Previously, we developed a bioluminescent detection method for the 0157 VT gene, which involved the luciferin-luciferase reaction following transformation of pyrophosphate produced during PCR to ATP by adenosine 5 phosphosulfate and ATP sulfiirylase. However, the sensitivity of this technique was insufficient due to slight light emission of APS during the luciferin-luciferase reaction, leading to elevation of the blank value. Furthermore, APS and ATP sulfiirylase, which are expensive, display poor stability for utility in routine analysis. 

DNA polymerase I, Klenow Fragment, EXO(-) was obtained from Funakoshi (Tokyo, Japan). Luciferase, Adenosine-5 -triphosphate sulfurylase, adenosine 5 -phosphosulfate sodium salt, D-Luciferin sodium salt, and magnesium acetate Tetra hydrate were obtained from Sigma (MO, USA). Deoxynucleotide and 2 -deoxyadenosine 5 -0-(l-thiotriphosphate) were obtained from Amersham Pharmacia Biotech (UK). Other chemicals were of an analytical-reagent grade. Instruments ... 

---