Coordination properties of 3′,5′-cyclic adenosine monophosphate towards copper(II) ions

The metal binding ability of 3′,5′-cyclic adenosine monophosphate (3′,5′-cAMP) molecule using copper(II) ion, as an example of biologically available divalent metal ion, was investigated by potentiometry, EPR and differential spectroscopy (UV-Vis, CD). One complex with stoichiometry Cu(3′,5′-cAMP)⁺ was found, where Cu(II) ion is bound by N-7 nitrogen of adenine moiety.     

Localization of 3′,5′-cyclic adenosine monophosphate phosphodiesterase (cAMP-PDEase) activity in isolated bovine thyroid plasma membranes

Summary Isolated bovine thyroid plasma membrane preparations were obtained by isopycnic density gradient centrifugution. Cyclic AMP-PDEase (EC 3.1.4.c) activity has been demonstrated by electron microscopic histochemistry on the unit membrane of isolated bovine thyroid cells. 3-isobutyl-1-methyl-xanthine (IBMX) produced partial inhibiton, while omission of the substrate revealed no reaction product deposition. These observations correlated well with biochemical studies that showed 0.4% of the total cAMP-PDEase activity to be present in the plasma membrane preparations. Kinetic analysis of cAMP hydrolysis yielded two apparent Michaelis constants for the homogenate and the plasma membrane-rich fraction. Dose-response curves for IBMX inhibition showed cAMP-PDEase of the homogenate to be more sensitive to inhibition than that of the plasma membrane-bound enzyme. Furthermore, wash experiments indicate that the plasma membrane-associated enzyme is tightly bound. This investigation strengthens our previous study and suggests that bovine thyroid cell plasma membranes contain a cAMP-PDEase that may be involved in interactions between the cell and the external environment in a manner yet to be determined.     

Effects of thyrotropin,prostaglandin E1 and iodide on cyclic 3′,5′-AMP concentration in dog thyroid slices

The kinetics and concentration effect on the relationship of thyrotropin (TSH) action on cyclic 3′,5′-AMP concentration has been studied in dog thyroid slices in vitro. TSH markedly increased cyclic 3′,5′-AMP level after 5 min, the effect reached a plateau after 10–60 min and slowly declined afterwards. TSH enhanced in parallel the cyclic 3′,5′-AMP level and the binding of iodide to proteins. For this latter effect of TSH, the four criteria of the validity of the Sutherland model for a hormonal action are therefore fulfilled. The effect of TSH on cyclic 3′,5′-AMP concentration in thyroid did not require the presence of a methylxanthine inhibitor of cyclic 3′,5′-AMP phosphodiesterase in the medium. Prostaglandin E1 increased cyclic 3′,5′-AMP levels in control and stimulated slices. The omission of Ca²⁺ in the incubation medium decreased the action of TSH but partial replacement of Na⁺ by K⁺ had little effect. Iodide, 1 μM to 100 μM, inhibited the action of TSH. This inhibitory effect was relieved by NaClO₄, methimazole and propylthiouracil (1 mM). The possible role of this inhibitory effect in an intracellular regulatory mechanism is discussed.     

Synthesis, spectroscopic, thermal and X-ray structure characterization of 1,3-propanediammonium tetrathiomolybdate and N,N,N′,N′-tetramethylethylenediammonium tetrathiomolybdate

The title compounds 1,3-propanediammonium tetrathiomolybdate, (1,3-pnH₂)[MoS₄], 1 and, N,N,N′,N′-tetramethylethylenediammonium tetrathiomolybdate, (tmenH₂)[MoS₄], 2, were prepared by reacting the ammonium salt of [MoS₄]²⁻ with the corresponding organic diamine. In 1 and 2 the organic diamines 1,3-propanediamine (1,3-pn) and N,N,N′,N′-tetramethylethylenediamine (tmen) are present in their diprotonated form. The reaction of 1 or 2 with [Ni(en)₃]Cl₂ · 2H₂O (en is ethylenediamine) results in the formation of the highly insoluble complex tris(ethylenediamine)Ni(II) tetrathiomolybdate, [Ni(en)₃][MoS₄], in quantitative yields. 1 and 2 have been characterized by chemical analysis, vibrational, UV-Vis and NMR spectroscopy, TG-DTA-MS and single crystal X-ray crystallography. Compound 1 is thermally more stable compared to 2. Both complexes decompose in a single step forming amorphous molybdenum sulfide. The structure of the title complexes can be described as consisting of tetrahedral [MoS₄]²⁻ dianions which accept a complex series of H-bonds from the organic dications. The strength and number of these hydrogen bonds affect the Mo-S bond lengths.     

Studies on the uptake and metabolism of adenosine 3':5'-cyclic monophosphate and N6,O2-dibutyryl 3':5'-cyclic adenosine monophosphate in sea urchin eggs

Structurally abnormal mitochondria were found in Candida utilis cells grown in the presence of either high copper concentrations or copper deficiency as compared with cells grown in standard media (copper 60 μg/l). In cells grown under conditions of copper deficiency the average size of the mitochondria is 0.6 μm compared with 0.2 μm of the normal cell, and the cristae have an abnormal appearance. In cells grown in the presence of high copper concentrations the mitochondrial size is up to 2–3 μm with poorly developed cristae and abnormal appearance of the matrix area.     

Localization of 3',5'-cyclic adenosine monophosphate phosphodiesterase (cAMP-PDEase) activity in isolated bovine thyroid plasma membranes

Isolated bovine thyroid plasma membrane preparations were obtained by isopycnic density gradient centrifugation. Cyclic AMP-PDEase (EC 3.1.4.c) activity has been demonstrated by electron microscopic histochemistry on the unit membrane of isolated bovine thyroid cells. 3-isobutyl-1-methyl-xanthine (IBMX) produced partial inhibition, while omission of the substrate revealed no reaction product deposition. These observations correlated well with biochemical studies that showed 0.4% of the total cAMP-PDEase activity to be present in the plasma membrane preparations. Kinetic analysis of cAMP hydrolysis yielded two apparent Michaelis constants for the homogenate and the plasma membrane-rich fraction. Dose-response curves for IBMX inhbition showed cAMP-PDEase of the homogenate to be more sensitive to inhibition than that of the plasma membrane-bound enzyme. Furthermore, wash experiments indicate that the plasma membrane-associated enzyme is tightly bound. This investigation strengthens our previous study and suggests that bovine thyroid cell plasma membranes contain a cAMP-PDEase that may be involved in interactions between the cell and the external environment in a manner yet to be determined.     

Enzymatic characteristics of two novel Myxococcus xanthus enzymes, PdeA and PdeB, displaying 3′,5′- and 2′,3′-cAMP phosphodiesterase, and phosphatase activities

Myxococcus xanthus PdeA and PdeB, enzymes homologous to class III 3′,5′-cyclic nucleotide phosphodiesterases, hydrolyzed 3′,5′- and 2′,3′-cyclic AMP (cAMP) to adenosine, and also demonstrated phosphatase activity toward nucleoside 5′-tri-, 5′-di-, 5′- and 3′-monophosphates with highest activities for nucleoside 5′-monophosphates. The substrate specificities of PdeA and PdeB show no similarity to that of any known cNMP phosphodiesterase, nucleotidase, or phosphatase. The enzyme activities of PdeA and PdeB were stimulated by 50 μM Mn²⁺ or Co²⁺. The K_m values of PdeA and PdeB for 3′,5′-cAMP, 2′,3′-cAMP, 5′-ATP, and 5′-AMP were in the low micromolar range (1.4-12.5  μM).     

Occurrence of adenosine 3′:5′-cyclic monophosphate in plant tissues

A procedure is described which unequivocally demonstrates the presence of adenosine 3′:5′-cyclic monophosphate in Phaseolus vulgaris. Its concentration was determined spectrophotometrically at 2·6–9·2 nmol g^?1 of tissue (dry wt) for 6-day-old seedlings and about one-tenth of this in 13-day-old plants.     

Exercise-induced increases in myocardial adenosine 3′,5′-cyclic monophosphate and phosphodiesterase activity

Numerous cellular biochemical events caused by hormones are mediated throught cyclic AMP. Although many changes occur in the cell during exercise that could be attributed to this nucleotide, little evidence is available implicating it as an important regulator of exercise metabolism. In this investigation it was found that a 60 min bout of treadmill exercise caused a 2.4-fold increase in myocardial cyclic AMP immediately following the work. Rather than the imemediate nucleotide hydrolysis that was expected, it was found that the elevated cyclic AMP level remained for approx. 24 h before returning to control levels. Cardiac glycogen fell to 30% of control after work but supercompensated 60% above control within 1 h following exercise. Therefore, cardiac cyclic AMP was elevated at a time when glycogen was being synthesized. Study of the temporal relationship between the exercise-induced increase in cyclic AMP and cyclic nucleotide phosphodiesterase indicated that the work caused an increase in the hearts' capacity to hydrolyze cyclic AMP. Measurement of heart phosphodiesterase at substrate concentrations of 1.0 and 100 μM produced significant increased in enzyme activity immediately following exercise which remained elevated for 48 h and was back to control activity 96 h following work. These data present a potentially fascinating model for the study of the dissociation between cyclic AMP, glycogenesis and elevations in phosphodiesterase activity in the heart.     

Effects of thyrotropin and N6,O2′-dibutyryl cyclic 3′,5′-adenosine monophosphate on prostaglandin levels in thyroid

We have shown that TSH increases PG levels in isolated bovine thyroid cells. We now report that TSH also increases PG levels in rat and mouse thyroid, and that these effects may be mediated via cyclic AMP. PG and cyclic AMP levels in intact rat and mouse thyroid lobes were measured by radioimmunoassay. During 60-min incubations at 37°C, 25 mU/ml TSH effected a 75–83% increase in PGE₁ and PGF_2α ”equivalents“ in rat thyroid; parallel measurements of endogenous cyclic AMP in these intact thyroid lobes revealed that maximal TSH-induced increase in cyclic AMP also required 60-min incubations. In mouse thyroid, 5 mU/ml TSH increased PGE₁ and PGF_2α levels 38–82% above basal; this TSH effect was evident within 15 min of incubation, thus mimicking the time-course of TSH-induced increase in mouse thyroid cyclic AMP. Exogenous DBcAMP, 0.5 to 3 mM, effected a dose-related increase in mouse thyroid PG levels. The stimulatory effects of both TSH and DBcAMP on mouse thyroid PG levels were abolished by aspirin and indomethacin. These studies suggest that TSH-induced increase in endogenous PG levels in thyroid may be mediated by cyclic AMP.     

Mass spectrometric identification of adenosine 3′:5′-cyclic monophosphate isolated from a higher plant tissue

Mass spectrometric evidence is presented confirming the identification of the adenosine nucleotide previously isolated from tissues of Phaseolus vulgaris as adenosine 3′: 5′-cyclic monophosphate.     

Identification of N, O-diacetyl-, N-acetyl- and N-glycolylneuraminyl-(2 → 3)-lactose in rat urine

The structure of three neuraminyl-oligosaccharides isolated from rat urine-have been studied by chromatographic and mass spectrometric analyses of different hydrolysis and methylation products. The structures of the oligosaccharides were identifies as O-α-N-acetyl(O-acetyl)neuraminyl-(2 → 3)-O-β-galactopyranosyl-(1 → 4)-glucopyranose, O-α-N-acetylneuraminyl-(2 → 3)-O-β-galactopyranosyl-(1 → 4)-glucopyranose and O-α-N-glycolylneuraminyl-(2 → 3)-O-β-galactopyranosyl-(1 → 4)-glucopyranose.     

Anion effects on ionogel formation in N,N′-dialkylimidazolium-based ionic liquids

Addition of an appropriate concentration of water to either 1-decyl-3-methylimidazolium bromide or its nitrate analog is shown to trigger the self-assembly of the ionic liquid (IL) and the concomitant formation of gel (“ionogel”) phases adopting a rich variety of structural motifs. Infrared and ¹H nuclear magnetic resonance studies indicate that water addition disrupts hydrogen bonding between the imidazolium ring and the IL anion, and that gelation involves the partial replacement of these bonds with H-bonds between the anion and water. Using small angle X-ray scattering, the relationship of the water content and the nature of the IL anion to the mesoscopic structure of the ionogels has been determined. Of particular note is the observation that by adjustment of the ionogel composition (water content or anion), near-monodomain materials can be formed with either lamellar, 2-D hexagonal or 3-D cubic structures with tunable lattice dimensions.     

Identification and characterization of the adenosine 3′,5′-cyclic monophosphate binding proteins appearing during the development of Dictyostelium discoideum

A photosensitive, radioactive analogue of cyclic adenosine monophosphate, 8-azido-adenosine 3′,5′-[³²P]monophosphate (8-N₃-cyclic AMP), was used to label the cyclic AMP binding proteins of Dictyostelium discoideum. During development cytosolic proteins appear which are specifically labeled by the photoaffinity agent. The proteins are developmentally regulated since they are only found in starved, developing cells. Unlabeled cyclic AMP competes specifically with the labeled analogue for protein binding sites in contrast to unlabeled 5′-AMP which does not compete. A mutant which develops spores but is deficient in stalk cell production produces a different set of cyclic AMP binding proteins from the parent strain.     

Uptake of adenosine 3',5'-cyclic monophosphate and isoproterenol by filarial parasites

The filarial parasites Litomosoides carinii and Dipetalonema viteae both show transcuticular uptake of adenosine 3',5'-cyclic monophosphate but isoproterenol is taken up by D. viteae only. The importance of this difference is discussed from the point of view of metabolic regulation. Inhibition of uptake by lectins indicates the involvement of surface sugar moieties in the transport processes.     

Guanosine 3′:5′-cyclic monophosphate changes during germination of Hordeum vulgare

Guanosine 3′: 5′-cyclic monophosphate (cGMP) isolated from barley seeds and seedlings was purified using neutral alumina and anion-exchange column chromatography, then descending paper chromatography, and finally estimated by means of radioimmunoassay. The putative compound was identified on cellulose chromatography in three solvent systems.During the early phase of the germination, the cGMP content decreased steadily from 30 fmol/g of dry seeds to undetectable amounts in seeds after 18 h of germination. The process of seedling growth was associated with the increase of cGMP concentration. Nine-day-old seedlings contained 147 and 200 fmol/g of fresh weight in the roots and in the coleoptiles plus leaves, respectively.