Nucleotide Synthesis. II. Nucleotide p-Nitrophenyl and 2,4-Dinitrophenyl Esters<sup>1,2</sup>

Borden, R K; Smith, Martha E.

doi:10.1021/jo01348a034

Cited by 27 publications

(13 citation statements)

References 1 publication

(3 reference statements)

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“…p -Nitrophenyl 5′-adenosine monophosphate ( p -Nph-5′-AMP) was synthesized in analogy to a described procedure (Borden and Smith, 1966 ; Ivanovskaya et al, 1987 ). Adenosine-5′-monophosphate disodium salt ( 17 , see Figure 4 , 1.0 g) was dissolved in 20 mL of deionized water.…”

Section: Methodsmentioning

confidence: 99%

Substrate-Dependence of Competitive Nucleotide Pyrophosphatase/Phosphodiesterase1 (NPP1) Inhibitors

et al. 2017

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Nucleotide pyrophosphatase/phosphodiesterase type 1 (NPP1) is a membrane glycoprotein involved in the hydrolysis of extracellular nucleotides. Its major substrate is ATP which is converted to AMP and diphosphate. NPP1 was proposed as a new therapeutic target in brain cancer and immuno-oncology. Several NPP1 inhibitors have been reported to date, most of which were evaluated vs. the artificial substrate p-nitrophenyl 5 ′ -thymidine monophosphate (p-Nph-5 ′ -TMP). Recently, we observed large discrepancies in inhibitory potencies for a class of competitive NPP1 inhibitors when tested vs. the artificial substrate p-Nph-5 ′ -TMP as compared to the natural substrate ATP. Therefore, the goal of the present study was to investigate whether inhibitors of human NPP1 generally display substrate-dependent inhibitory potency. Systematic evaluation of nucleotidic as well as non-nucleotidic NPP1 inhibitors revealed significant differences in determined K i values for competitive, but not for non-and un-competitive inhibitors when tested vs. the frequently used artificial substrate p-Nph-5 ′ -TMP as compared to ATP. Allosteric modulation of NPP1 by p-Nph-5 ′ -TMP may explain these discrepancies. Results obtained using the AMP derivative p-nitrophenyl 5 ′ -adenosine monophosphate (p-Nph-5 ′ -AMP) as an alternative artificial substrate correlated much better with those employing the natural substrate ATP.

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Section: Methodsmentioning

confidence: 99%

Substrate-Dependence of Competitive Nucleotide Pyrophosphatase/Phosphodiesterase1 (NPP1) Inhibitors

et al. 2017

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“…The reaction of a nucleoside phosphate with 2,4-dinitrofluorobenzene provides a convenient route to nucleoside phosphorofluoridates (7). However, 2,4-dinitrofluorobenzene reacts with a variety of nucleophiles 4 5.00 class of unwanted product in reaction with nucleotides (8).…”

Section: Discussionmentioning

confidence: 99%

Reaction of a nucleoside 2,4-dinitrophenyl phosphate with fluoride; A convenient method for the preparation of the nucleoside phosphorfluoridate

1975

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“…Several preliminary attempts were made to determine whether endogenous cyclic AMP could be detected in barley seed extracts using the binding protein assay described by Gilman (7). This assay can detect 0.1 pmole of cyclic AMP and has been used to measure amounts of cyclic AMP (1)(2)(3)(4)(5)(6)(7)(8)(9)(10)(11)(12)(13)(14)(15)(16)(17)(18) pmoles/mg tissue) present in animal tissues (7). Barley seed extracts were found to contain a factor (or factors) which inhibit the binding of 'H-cyclic AMP to protein kinase and which disappears after incubation with cyclic 3':5'-nucleotide phosphodiesterase (100 jug/ml) for 3 hr at 30 C. However, the inhibitory material is equivalent to only 0.1 to 0.2 pmoles of cyclic AMP/mg tissue-values which are close to the limit of detection of the assay.…”

Section: Methodsmentioning

confidence: 99%

“…1). Accordingly, an authentic sample of the ammonium salt of AMP-F was prepared by reaction of 5'-AMP with 2, 4-dinitrofluorobenzene as previously described (2,29).…”

Section: Methodsmentioning

confidence: 99%

Formation of Adenosine 5′-Phosphorofluoridate and the Assay of Adenyl Cyclase in Barley Seeds

Alvarez¹,

Moore

Vandepeute

1974

Plant Physiol.

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Barley seed (Hordeum vulgare L.) homogenates contain an apparent enzymatic activity which catalyzes the synthesis of adenosine 5'-phosphorofluoridate from magnesium-adenosine 5'-triphosphate and sodium fluoride. Formation of this compound may interfere with some adenyl cyclase assays which use fluoride as a component of the incubation medium. Neither adenyl cyclase activity nor endogenous adenosine 3': 5'-monophosphate was detected in barley seed homogenates or extracts.The mechanism of action of some animal hormones involves an alteration in the intracellular level of adenosine 3': 5'-monophosphate (cyclic AMP) (9,21,22,26). These hormones stimulate the activity of a membrane-bound adenyl cyclase system and thereby increase the enzymatic production of cyclic AMP from ATP. Sodium fluoride markedly stimulates adenyl cyclase activity in broken cell preparations from a variety of animal tissues (21,22), and it is routinely used in the assay of adenyl cyclase (11). A speculative explanation for the mechanism of fluoride action has been proposed which involves the formation of adenosine 5'-phosphorofluoridate as a reactive intermediate (1).Recently, preliminary evidence for the presence of adenyl cyclase (30) and endogenous cyclic AMP (16) in plant tissues has been presented. The synthesis of a cyclic AMP-like compound in plant tissues incubated with "4C-adenine has also been observed (15,18,23). Several reports indicate that the cyclic nucleotide promotes the secretion of some hydrolytic enzymes from embryoless barley seeds (4-6, 17, 19, 24 MATERIALS AND METHODSAdenyl Cyclase Assay. Adenyl cyclase activity was assayed by the method of Ramachandran (20). Tissues were homogenized in 10 volumes of cold 10 mm tris, pH 7.5, containing 0.25 M sucrose. Assays were performed for 15 min at 30 C in the presence or absence of 10 mm sodium fluoride. Washed particles from guinea pig hearts were prepared according to the method of Drummond and Duncan (3). The incubation medium for the adenyl cyclase assay was identical to that described previously (3), except for the concentrations of ATP (6 mM) and magnesium sulfate (9 mM).Cyclic AMP Assays. Endogenous cyclic AMP was assayed by the methods of Wastila et al. (28) and Gilman (7). Barley seed extracts were prepared in accordance with the method of Gilman (7).Apparent Enzymatic Formation of AMP-F. The synthesis of AMP-F was measured by a modification of an assay procedure described by Krishna et al. (11) for the determination of adenyl cyclase activity. Seeds were powdered and homogenized in 10 volumes of cold 10 mm MES buffer (pH 6.0), 1 mM dithiothreitol, and 10% (w/v) sucrose. The mixture was centrifuged at 30,000g for 10 min, and the supernatant fraction was used directly in the assay. The incubation medium consisted of 40 mm MES (pH 6.0), 3.3 mm magnesium sulfate, 10 mM caffeine, 0.1 mm ATP (0.5 fic "4C-ATP-34 mc/mmole), 0.1 M sodium fluoride and 0.18 ml of the enzyme preparation in a final volume of 0.6 ml. After an incubation period of 60 min at 30 C, the solutions ...

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Nucleotide Synthesis. II. Nucleotide p-Nitrophenyl and 2,4-Dinitrophenyl Esters^1,2

Cited by 27 publications

References 1 publication

Substrate-Dependence of Competitive Nucleotide Pyrophosphatase/Phosphodiesterase1 (NPP1) Inhibitors

Substrate-Dependence of Competitive Nucleotide Pyrophosphatase/Phosphodiesterase1 (NPP1) Inhibitors

Reaction of a nucleoside 2,4-dinitrophenyl phosphate with fluoride; A convenient method for the preparation of the nucleoside phosphorfluoridate

Formation of Adenosine 5′-Phosphorofluoridate and the Assay of Adenyl Cyclase in Barley Seeds

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Nucleotide Synthesis. II. Nucleotide p-Nitrophenyl and 2,4-Dinitrophenyl Esters1,2

Cited by 27 publications

References 1 publication

Substrate-Dependence of Competitive Nucleotide Pyrophosphatase/Phosphodiesterase1 (NPP1) Inhibitors

Substrate-Dependence of Competitive Nucleotide Pyrophosphatase/Phosphodiesterase1 (NPP1) Inhibitors

Reaction of a nucleoside 2,4-dinitrophenyl phosphate with fluoride; A convenient method for the preparation of the nucleoside phosphorfluoridate

Formation of Adenosine 5′-Phosphorofluoridate and the Assay of Adenyl Cyclase in Barley Seeds

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Nucleotide Synthesis. II. Nucleotide p-Nitrophenyl and 2,4-Dinitrophenyl Esters^1,2