Role of cyclic adenosine-3′,5′-monophosphate in olfactory reception

The action of cyclic adenosine-3,5-monophosphate (3,5-AMP) and of substances modifying the rate of its breakdown (inhibitors and activators of phosphodiesterase) on the olfactory epithelium was investigated in frogs. The slow electrical response of the olfactory epithelium to stimulation by solutions of various substances was recorded. Cyclic 3,5-AMP and its dibutyryl derivative were found to excite the olfactory receptors effectively. Responses to these substances developed after an appreciably longer delay than responses to stimulation by solutions of odiferous substances. It is postulated that the depolarizing action of 3,5-AMP and dibutyryl 3,5-AMP is manifested only after they have penetrated inside the receptor cell through its membrane. Both 5-AMP and cyclic 2,3-AMP were ineffective. In the next series of experiments the integral receptor potential was recorded in response to short stimulation by the vapor of an odiferous substance. The duration of this potential was increased after treatment of the olfactory epithelium with phosphodiesterase inhibitors: methylxanthines or papaverine. Conversely, the negative wave of the integral receptor potential was shortened under the influence of the phosphodiesterase activator imidazole. Cyclic 3,5-AMP is considered to play the role of mediator in the mechanism of excitation of the olfactory receptor; during interaction between an odiferous substance and the receptor, adenyl cyclase is activated and the concentration of 3,5-AMP increases; this, in turn, causes depolarization of the receptor cell membrane.Institute of Chemical Physics, Academy of Sciences of the USSR, Moscow. Translated from Neirofiziologiya, Vol. 5, No. 4, pp. 415–422, July–August, 1973.     

Studies of kidney plasma membrane adenosine-3′,5′-monophosphate-dependent protein kinase

Porcine kidney cortex was utilized for the preparation of plasma-membrane-enriched and soluble cytoplasmic (cytosol) fractions for the purpose of examining the relative properties of cyclic [³H]AMP receptor and cyclic AMP-dependent protein kinase activities of these preparations. The affinity, specificity and reversibility of cyclic [³H]AMP interaction with renal membrane and cytosol binding sites were indicative of physiological receptors.Binding sites of cytosol and deoxycholate-solubilized membranes were half-saturated at approx. 50nM and 100 nM cyclic [³H]AMP. Native plasma membranes exhibited multiple binding sites which were not saturated up to 1 mM cyclic [³H]AMP. Modification of the cyclic phosphate configuration or 2′-hydroxyl of the ribose moiety of cyclic AMP produced a marked reduction in the effectiveness of the cyclic AMP analogue as a competitor with cyclic [³H]AMP for renal receptors. The cyclic [³H]AMP interaction with membrane and cytosol fractions was reversible and the rate and extent of dissociation of bound cyclic [³H]AMP was temperature dependent. With the plasma-membrane preparation, dissociation of cyclic [³H]AMP was enhanced by ATP or AMP.Assay of both kidney subcellular fractions for protein kinase activity revealed that cyclic AMP enhanced the phosphorylation of protamine, lysine-rich and arginine-rich histones but not casein. The potency and efficacy of activation of renal membrane and cytosol protein kinase by cyclic AMP analogues such as N⁶-butyryl-adenosine cyclic 3′,5′-monophosphate or N⁶,O²-dibutyryl-adenosine cyclic 3′,5′-monophosphate supported the observations on the effectiveness of cyclic AMP analogues as competitors with cyclic [³H]AMP in competitive binding assays.This study suggested that the membrane cyclic [³H]AMP receptors may be closely associated with the membrane-bound catalytic moiety of the cyclic AMP-dependent protein kinase system of porcine kidney.     

A general method for visualizing enzymes releasing adenosine or adenosine-5′-monophosphate

Biochemical Genetics -     

N,N′-carbonyldiimidazole-induced diketopiperazine formation in aqueous solution in the presence of adenosine-5′-monophosphate

Summary 3(2)-O-glycyl-adenosine-5-monophosphate is an intermediate in the conversion of N-[imidazolyl-(1)-carbonyl]-glycine to diketopiperazine in the presence of adenosine-5-monophosphate. The significance of these observations to prebiotic chemistry is discussed.Abbreviations AMP adenosine-5-monophosphate - A adenosine     

The 5′-Nor Aristeromycin Analogues of 5′-Deoxy-5′-Methylthioadenosine and 5′-Deoxy-5′-Thiophenyladenosine

To extend the potential of 5′-noraristeromycin (and its enantiomer) as potential antiviral candidates, the enantiomers of the carbocyclic 5′-nor derivatives of 5′-methylthio-5′-deoxyadenosine and 5′-phenylthio-5′-deoxyadenosine have been synthesized and evaluated. None of the compounds showed meaningful antiviral activity.     

Measurement of mitochondrial oxidative phosphorylation: Selective inhibition of adenylate kinase activity by P1,P5-di-(adenosine-5′)-pentaphosphate

A biochemical assay for the measurement of ATP synthesis coupled to electron transport in the presence of adenylate kinase was developed as an alternative to using the conventional Clark-type oxygen electrode. The assay utilizes P¹,P⁵-di-(adenosine-5′)-pentaphosphate which is shown to be a competitive inhibitor with MgADP for rat liver mitochondrial adenylate kinase (K_i = 7.04 × 10^?8m) and was found to have no effect on oxidative phosphorylation of either intact mitochondria or submitochondrial particles.     

Identification and quantitation of adenosine-3′:5′-cyclic monophosphate in plants using gas chromatography-mass spectrometry and high-performance liquid chromatography

Direct evidence has been obtained for the presence of adenosine-3:5-cyclic monophosphate (cAMP) in tobacco (Nicotiana tabacum L.) callus tissue cultures, bean (Phaseolus vulgaris L.) seedlings and immature kernels of sweet corn (Zea mays L.) through the use of a highly specific and sensitive gas chromatography-mass spectrometric assay. Levels of endogenous cAMP ranged from 70 to 126 pmol/g fresh weight. Corresponding levels of cAMP determined for the same samples using radioimmunoassay were consistently three to four times higher. Contrary to previous reports for citrus plants, measurable levels of cAMP could not be detected in young lemon leaves within the limits of detection of the mass-spectrometric assay method. In the case of tobacco callus tissue, the coumarin glucoside, scopolin, which was present in large amounts and showed similar chromatographic behaviour to cAMP, interferred strongly with the mass-spectrometric measurements of cAMP in inadequately purified extracts. The use of high-performance liquid chromatography, in addition to standard chromatographic purification methods, produced highly purified plant extracts for quantitation of cAMP and also provided a method for the separation of cAMP from its 2:3-isomer.Abbreviations cAMP adenosine-3:5-cyclic monophosphate - 2:3-cAMP adenosine-2:3-cyclic monophosphate - GC-MS-MID combined gas chromatography-mass spectrometry with selected multiple-ion-detection - HPLC high-performance liquid chromatography - RIA radioimmunoassay - TLC thin-layer chromatography     

Regulation of adenosine-5′-phosphosulfate sulfotransferase activity by H2S in Lemna minor L.

When Lemna minor L. is transferred to an atmosphere with H₂S, there is a rapid loss of extractable adenosine-5-phosphosulfate sulfotransferase activity. The activity is restored within 24 h in an atmosphere without H₂S. This restoration of activity is completely inhibited by cycloheximid but not by chloramphenicol. In vitro, S^2- up to 5 mM and cysteine, methionine, and glutathione up to 50 mM do not inhibit the enzyme. The activities of ATP sulfurylase and O-acetyl-L-serine sulfhydrylase are not affected significantly by H₂S. The physiological significance of the regulation of adenosine-5-phosphosulfate sulfotransferase is discussed.Abbreviations APS adenosine-5-phosphosulfate - PAPS adenosine-3-phosphate-5-phosphosulfate - BSA bovine serum albumin - DTT dithiothreitol - POPOP 1,4-di [2-(5-phenyloxazolyl)]-benzene - PPO 2,5-diphenyloxazol This is no. 6 in the series Regulation of Sulfate Assmilation in Plants     

Involvement of cyclic adenosine-3′, 5′-monophosphate in chloronema differentiation in protonema cultures of Funaria hygrometrica

The role of purine and pyrimidine ribosides, nucleotides and substituted xanthines in the differentiation of chloronema filaments in suspension cultures of protonema of the moss Funaria hygrometrica Hedw. has been examined. Cyclic adenosine-3,5-monophosphate (cAMP) and mono-and dibutyryl cAMP evoked the maximum response in wild-type protonema. ADP and ATP also enhanced chloronema differentiation but were less active than cAMP; pyrimidine derivatives were completely inactive. Inhibitors of cyclic-nucleotide phosphodiesterase aminophylline, theophylline and ICI 58, 301 (3-acetamido-6-methyl-8-n-propyl-s-triazolo-(4,3a)-pyrazine)-mimicked the effect of cAMP. A leaky, chloronema-repressed mutant was isolated and in this mutant cAMP was much more active than cyclic guanosine monophosphate and ADP in enhancing chloronema differentiation. These results strongly indicate that cAMP is involved in chloronema differentiation in Funaria, and a hypothesis on growth regulation in protonema cell cultures is proposed.Abbreviations cAMP, cyclic AMP cyclic adenosine-3, 5-monophosphate - cCMP, cGMP, cIMP cyclic cytosine-, guanosine-and inosine-3, 5-monophosphates, respectively - IAA indole-3-acetic acid - ICI 58,301 3-acetamido-6-methyl-8-n-propyl-s-triazolo-(4,3a)-pyrazine     

Synthesis and evaluation of new N6-substituted adenosine-5′-N-methylcarboxamides as A3 adenosine receptor agonists

A number of N⁶-substituted adenosine-5′-N-methylcarboxamides were synthesised and their pharmacology, in terms of their receptor affinity, selectivity and cardioprotective effects, were explored. The first series of compounds, 4a–4f and 5a–5f, showed modest receptor affinity for the A₃AR with K_i values in the low to mid μM range. However, the incorporation of a 4-(2-aminoethyl)-2,6-di-tert-butylphenol group in the N⁶-position (in compounds 4g and 5g) significantly improved the affinity with K_i values of 30 and 9 nM, respectively. Improvements in affinity, as well as selectivity were seen when a functionalised linker was introduced. The N⁶-phenyl series, compounds 7a–7d, demonstrated low to mid nanomolar receptor affinities (K_i = 2.3–45.0 nM), with 7b displaying 109-fold selectivity for the A₃AR (vs A₁). The N⁶-benzyl series 9a–9c also proved to be potent and selective A₃AR agonists and the longer chain length linker 13 was tolerated at the A₃AR without abrogation of affinity or selectivity. Cardioprotection was demonstrated by a simulated ischaemia cell culture assay, whereby 7b, 7c, 9a, 9b and 9c all showed cardioprotective effects at 100 nM comparable or better than the benchmark A₃AR agonist IB-MECA, but which were indistinguishable by statistical analysis. For example, compound 9c reduced cell death by 68.0 ± 3.6%.     

MECHANISM OF INACTIVATION OF S-ADENOSYL-L-HOMOCYSTEINE HYDROLASE BY 5′-DEOXY-5′-S-ALLENYLTHIOADENOSINE AND 5′-DEOXY-5′-S-PROPYNYLTHIOADENOSINE

5′-Deoxy-5′-S-allenylthioadenosine 1 and 5′-deoxy-5′-S-propnylthioadenosine 2, derived from adenosine, were prepared. 1 and 2 caused irreversible inactivation of AdoHcy hydrolase. ESI mass spectra analysis of the inactivated enzyme demonstrated that 1 and 2 were type II “mechanism-based” inhibitors.     

Synthesis of 5′-Fluoro-5′-deoxy-and 5′-Amino-5′-Deoxytoyocamycin and Sangivamycin and Some Related Derivatives

Abstract

A series of 5′-substituted analogs of toyocamycin were prepared by condensation of silylated 4-amino-6-bromo-5-cyanopyrrolo[2,3-d]pyrimidine with protected 5-azido-5-deoxy- or 5-fluoro-5-deoxyribofuranose followed by debromination and deblocking. Alternatively, 5′-azido-5′-deoxytoyocamycin was prepared by azidation of toyocamycin. Conversion of the 5-nitrile function of the toyocamycin derivatives into a carboxamide or a thiocarboxamide gave the corresponding analogs of sangivamycin or thiosangivamycin while reduction of the 5′-azido-5′-deoxy nucleosides provided 5′-amino-5′-deoxy derivatives.     

5′-nucleotidase

Summary The distribution of 5-nucleotidase activity in rat liver shows a sexual dependence. In male liver the activity in the bile canalicular wall is most pronounced, whereas the activity at the sinusoidal border of the liver parenchymal cell is slightly more in the female rat. Castration and treatment with sex hormones change the distribution pattern. The greatest variations in enzyme activity are seen at the bile canalicular site of the liver cell. These changes are probably an expression of the altered functional state of the liver cell.     

5′-Nucleotidase

Summary The distribution of 5-nucleotidase in several tissues of rat and mouse has been investigated. The enzyme is greatly specific in dephosphorylating 5-nucleotides. Two 5-nucleotidases seem to exist: one showing greatest activity at pH 5.0, while the other is most active at pH 7.0–7.5. The localization of these two enzymes is not identical. At the acid pH the deoxyribonucleotides are dephosphorylated faster than the ribonucleotides, while at the neutral pH the ribonucleotides are hydrolysed more rapidly. In contrast to the non-specific phosphatases the nucleotidases can be stimulated by magnesium and manganese ions. The acid nucleotidase can be considered as a lysosomal enzyme. The neutral nucleotidase can be involved in transport processes in capillaries and sinusoids. Other localizations of the enzyme suggest a role of the enzyme in the catabolism of nucleic acids.     

5′-Nucleotidase

Summary To get more insight in the function of 5-nucleotidase catabolic and anabholic processes were investigated in which 5-nucleotides are involved. The catabolism of adenosine-5-monophosphate was studied by investigating the reaction products obtained after incubation of homogenates of several organs of rat and mouse with adenosine-5-monophosphate and with adenosine. Two experimental tumours of the mouse were investigated in the same way. It was found that in tissues containing a high activity of 5-nucleotidase other enzymes involved in the catabolism of 5-nucleotides, such as nucleosidase, adenosine deaminase and adenosine-5-monophosphate deaminase could also be demonstrated.The anabolic processes in which 5-nucleotides are involved had been studied by investigating the incorporation of tritium-labeled thymidine in several tissues of the mouse. It appeared that in cells showing a high 5-nucleotidase activity no incorporation of radioactive thymidine could be found, while in cells showing incorporation of thymidine enzyme activity could not be demonstrated.A discussion is given about the possible role of 5-nucleotidase in the control of nucleic acid biosynthesis and in the catabolism of nucleic acids.Abbreviations used DNA deoxyribonucleic acid - RNA ribonucleic acid - AMP Adenosine-5-monophosphate - ADP Adenosine-5-diphosphate - ATP Adenosine-5-triphosphate - IMP Inosine-5-monophosphate - GMP Guanosine-5-monophosphate - GDP Guanosine-5-diphosphate - GTP guanosine-5-triphosphate - CMP Cytidine-5-monophosphate - CDP Cytidine-5-diphosphate - CTP Cytidine-5-triphosphate - UMP Uridine-5-monophosphate - UDP Uridine-5-diphosphate - UTP Uridine-5-triphosphate - TMP Thymidine-5-monophosphate - TDP Thymidine-5-diphosphate - TTP Thymidine-5-triphosphate - Ado Adenosine - Ad Adenine - Ino Inosine - Hypox Hypoxanthine - Xanth Xanthine - Xantho Xanthosine - Guano Guanosine - Gua Guanine - Ura Uracil - U Uridine - Cyt Cytidine - Cyto Cytosine - Thym Thymidine The corresponding deoxy-compounds have been indicated with the prefix d for instance dCMP, deoxycytidine-5-monophosphate.