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1.
The cyclic 35-nucleotide phosphodiesterase D3 was purified from Sinorhizobium fredii MAR-1. The native enzyme had a molecular weight of approximately 44.5kDa and a subunit molecular weight of approximately 21kDa as judged by SDS-gel electrophoresis. The pH optimum of the enzyme for the hydrolysis of cyclic AMP was approximately 6.0 with both acetate and Tris-maleate buffers. The optimum temperature for hydrolysing cyclic AMP was approximately 50C. No metal ion was required for activity and EDTA up to 2.5mM did not markedly affect the enzyme. However, methylxanthines, adenine and adenosine as well as 5-AMP, ATP, ADP and metal ions like Zn2+, Fe2+, Pb2+, Al3+ and Fe3+, were strongly inhibitory at 2.5mM.The D3 enzyme could hydrolyse both cyclic AMP and cyclic GMP with the apparent K m for cyclic AMP of approximately 0.23M.  相似文献   

2.
Summary Inhibition of growth of PY815 mouse mastocytoma cells in vitro by N6, O2-dibutyryladenosine 3,5 cyclic monophosphate (DB cyclic AMP) was accompanied by increases in intracellular cyclic AMP and histamine and minor changes in cytosolic cyclic AMP-dependent histone kinase activity. However, DEAE-cellulose chromatography revealed substantial changes in the relative proportions of the principal cyclic AMP-dependent protein kinases and in free cyclic AMP-binding protein after DB cyclic AMP treatment. The activity of cytosolic cyclic AMP-dependent protein kinase type I (PKI) decreased relative to cyclic AMP-dependent protein kinase type II (PKII) and there was an increase in a cytosol cyclic AMP-binding protein with little associated protein kinase activity. The relative changes in activity of PKI, PKII and cyclic AMP binding protein after DB cyclic AMP treatment may reflect events important in the regulation of growth and differentiation of mast cells.Abbreviations DB cyclic AMP N6,O2-dibutyryladenosine-3, 5-cyclic monophosphate - cyclic AMP adenosine 3,5-cyclic monophosphate - PKI type I cyclic AMP-dependent protein kinase - PKII type II cyclic AMP-dependent protein kinase  相似文献   

3.
The Ca2+ channel 1B subunit is a pore-forming component capable of generating N-type Ca2+ channel activity. Although N-type Ca2+ channel plays a role in a variety of neuronal functions, 1B-deficient mice exhibit normal life span without apparent abnormalities of behavior, histology or plasma norepinephrine level, presumably owing to compensation by some other Ca2+ channel 1 or subunit. In this study, we studied the levels of 1A, 1C, 1D, 1E, 1, 2, 3 and 4 mRNAs in adrenal gland of 1B-deficient mice. The 1A mRNA in homozygous mice was expressed at higher level than in wild or heterozygous mice, but no difference in the expression levels of 1C, 1D, 1E, 1, 2, 3 and 4 was found among wild, heterozygous and homozygous mice. The protein level of 1A in homozygous mice was also expressed at higher level than in wild or heterozygous mice. To examine whether increased expression is induced by cis-regulatory element within 5-upstream region of 1A gene, we examined lacZ expression in 1B-deficient × 1A6.3-lacZ mice (carrying a 6.3-kb 5-upstream fragment of 1A gene fused to E. coli lacZ reporter gene), which express lacZ in medullar chromaffin cells, but not in cortex. The levels of lacZ expression in homozygous 1B-deficient × 1A6.3-lacZ mice were higher than in wild or heterozygous mice. Therefore, a possible explanation of the normal behavior and plasma norepinephrine level of 1B-deficient mice is that compensation by 1A subunit occurs and that 6.3-kb 5-upstream region of 1A gene contains enhancer cis-element(s) for compensation in adrenal medulla chromaffin cells. (Mol Cell Biochem 271: 91–99, 2005)  相似文献   

4.
Summary Oligouridylates with more than eight chain units can serve as a template for the template-directed condensation of ImpA catalyzed by Pb2+ ion. The templates and the Pb2+ ion catalyst facilitate the formation of longer oligoadenylates with five or more units. The ratio of 3–5 linked oligomers to the 2–5 isomers increases with increasing chain length of the oligouridylate template. Short oligouridylates up to a hexamer tend to decrease the yield of oligoadenylates, and do not affect the selectivity of internucleotide linkage.Abbreviations EDTA ethylenediaminetetracetic acid - Tris tris(hydroxymethyl)aminomethane - A adenosine - ImpA adenosine 5-phosphorimidazolide - pA adenosine 5-phosphate - Ap adenosine 2(3)-phosphate - poly A polyadenylic acid - AppA P1,P2-diadenosine 5-diphosphate - pAp adenosine 2(3),5-diphosphate - ApA adenylyl adenosine - (pA)n (n = 2,3,) oligomers of pA - ImpApA 5-phosphorimidazolide of ApA - U uridine - pU uridine 5-phosphate - Up uridine 2(3)-phosphate - poly U polyuridylic acid - pUp uridine 2(3),5-diphosphate - (pU)n (n = 2,3,) oligomers of pU - (pU)n – (pA)m cooligomers composed of (pU)n and (pA)m units - AppUpUpUpUp pyrophosphate derived from pA and (pU)4 - AppUp P1-(adenosine 5)-P2-(uridine 2(3)-phosphate 5) -pyrophosphate - BAP bacterial alkaline phosphatase - VPD venom phosphodiesterase - N.P1 nuclease P1 - RNase A pancreatic ribonuclease - A* radioactive adenosine  相似文献   

5.
Many genes have been located in wheat chromosomes, yet little is known about the location of genes for resistance to Ustilago tritici, which causes loose smut. Crosses were made between the loose smut susceptible alien substitution lines Cadet 6Ag(6A) and Rescue 6Ag(6A) (lines in which Agropyron chromosome 6 is substituted by wheat chromosome 6A) and four cultivars resistant to U. tritici race T19: Cadet, Kota, Thatcher and TD18. The segregating progeny were tested for reaction to race T19 and for the level of binding with a monoclonal antibody specific to a chromosome 6A-coded seed protein. The antibody, which does not bind to seed protein extracts in the absence of the 6A chromosome, was used as a chromosome marker. An association was established between resistance to race T19 and the presence of chromosome 6A for each of the cultivars tested, indicating that resistance to race T19 resides in chromosome 6A. Ustilago tritici race T19 resistance in Cadet appears to be located in the short arm of chromosome 6A, based on the evaluation of the Cadet 6A long ditelosomic stock, which was susceptible, and the Cadet 6A-short: 6-Agropyron- short alien translocation stock, which was resistant.  相似文献   

6.
Synaptosome-enriched preparations from rat and guinea pig brain tissue vigorously accumulated [3H]-adenosine 5-monophosphate ([3H]AMP). When the accumulation of [3H]AMP was determined using incubation periods of 30 s or less, high concentrations of adenosine, dipyridamole and soluflazine did not inhibit the accumulation of label appreciably. The accumulation of [3H]AMP was saturable, temperature-dependent, osmotic-sensitive and exhibited structural specificity. Based on the kinetics of uptake by different subcellular fractions, and the inhibitory effects of other nucleotides, the uptake of AMP appeared to be mediated by three saturable systems with Kt values of approximately 0.2, 6, and 100 M. The transport system with the highest affinity for AMP was selectively inhibited by guanosine 5-monophosphate, and its Vmax was several fold higher in a myelin-enriched fraction than in synaptosome-enriched fractions. The transport system with the Kt6 M was selectively inhibited by ,-methylene adenosine diphosphate, and its Vmax was several times higher in a fraction enriched in high-density synaptosomes than in fractions enriched in low-density synaptosomes or myelin. Both of these transport systems were potently inhibited by ATP and ADP. Nucleotides that were either weak or inactive as inhibitors of AMP transport included 3-AMP, cyclic AMP, guanosine 5-diphosphate, and the 5-mononucleotides of cytosine, inosine, and uridine. GTP consistently enhanced uptake at concentrations 1 M. The transport of AMP was not Na+-dependent and was not inhibited by membrane depolarization. This transport system may mediate the release of AMP for subsequent conversion to adenosine extracellularly.Abbreviations used HEPES N-2-hydroxyethylpiperazine-N-2-ethanesulfonic acid - NBTI nitrobenzylthioinosine - ,-MeADP ,-methylene adenosine diphosphate - GTPgS guanosine-5-O-(3-thiotriphosphate) Special issue dedicated to Dr. Morris H. Aprison.  相似文献   

7.
Summary Evaporation of a solution of thymidine plus either theexo or theendo diastereomer of uridine cyclic 2,3-O, O-phosphorothioate (U > p(S) in 1,2-diaminoethane hydrochloride buffer gave the 2,5 and 3,5 isomers of (P-thio) uridylylthymidine (Up(S)dT) in a ratio of 1:2 with a combined yield of about 20%. These isomers were re-converted to U > p(S) and dT by a reaction that is known to proceed by an in-line mechanism. Both the 2,5 and 3,5 isomers gave as product the same diastereomer of U > p(S) that had been used originally in their formation. These dry-state prebiotic reactions (Verlander, Lohrmann, and Orgel 1973) are thus shown to be stereospecific, and both the 2,5 and 3,5 internucleotide bonds are formed by an in-line mechanism.Abbreviations DAE 1,2-diaminoethane - HPLC high pressure liquid chromatography - RNase bovine pancreatic ribonuclease A, EC 3.1.4.22 - TEAB triethylammonium bicarbonate - tris tris(hydroxymethyl)aminomethane - UMP(S) uridine monophosphorothioate - U > p uridine cyclic 2,3-phosphate - U > p(S) uridine cyclic 2,3-O, O-phosphorothioate - Up(S)dT (P-thio)uridylylthymidine - U2p(Rp-S)5dT (P-thio)uridylylthymidine with theR configuration at phosphorous, and a 2,5 internucleotide linkage  相似文献   

8.
Goat prothymosin , a highly acidic polypeptide of pl 3.5, 109 amino acid residues, has been isolated from lymphoid and non-lymphoid tissues of young female goats. Unlike rat, murine and porcine prothymosins , goat prothymosin appears at a higher concentration in the spleen compared with the thymus. The sequence of segments of the polypeptide involving known mutations has been determined, by automatic sequencing of its tryptic peptide fragments. The acidic amino acid-rich segment in the middle of the molecule, including residues 49–83, has not been sequenced. Goat prothymosin closely resembles bovine prothymosin , with only one substitution, proline for alanine at position 85. It also resembles human prothymosin , with only three substitutions. It differs more significantly from rat and murine prothymosins , by two deletions and three substitutions. The results show the highly conserved nature of the molecule, with substitutions at given positions only.Abbreviations ProT Prothymosin - T1 Thymosin 1 - MLR Mixed Lymphocyte Response - HPLC High Performance Liquid Chromatography - RIA Radioimmunoassay - B Aspartic acid or Asparagine - Z Glutamic acid or Glutamine  相似文献   

9.
A yeast-mycelium (Y-M) transition in Candida albicans was induced by exogenous yeast extract, adenosine, adenosine 5-monophosphate (AMP), adenosine 5-diphosphate (ADP), adenosine 35 cyclic monophosphate (cAMP) and its analogue N6, O2-dibutyryl adenosine 35-cyclic monophosphate (dbcAMP) in defined liquid medium at 25°C. Adenosine 5-triphosphate (ATP) was found to delay germ tube formation in yeast cells, whereas the cAMP phosphodiesterase inhibitors, theophylline and caffeine, induced a Y-M transition. Intracellular and extracellular cyclic AMP levels increased during the yeast-mycelium transition and maximum levels of intracellular cyclic AMP coincided with maximum germ tube formation. Of the many inducers and inhibitors of germ tube and mycelium formation in C. albicans tested, including incubation at 37°C or in the presence of 1.5mM CaCl2, the calmodulin inhibitor calmidazolium (R24571) added together with CaCl2 induced the highest intra- and extracellular cyclic AMP levels. These results confirm the involvement of cyclic AMP in the yeast-mycelium transition of C. albicans.  相似文献   

10.
Summary Erythrocyte soluble phosphodiesterase activities show at least two interconvertible states: aggregated and non-aggregated. In the former state the existence of a high affinity component for cyclic AMP is favoured and the system is more active with cyclic AMP than with cyclic GMP as substrate. When the system is in the non-aggregated state, no activity with cyclic GMP is detected. Conversion to the non-aggregated state is enhanced by dilution or by addition of magnesium ions.Upon isoelectric focusing of erythrocyte soluble extracts, phosphodiesterase activities can be resolved in two peaks: one specific for cyclic AMP located at pH 4.66 and the other specific for cyclic GMP focused at pH 4.95. Evidence would indicate that aggregation affects the enzyme specific for cyclic AMP.  相似文献   

11.
Summary Adenosine monophosphates (AMPs) cause the induction of floral buds in Impatients balsamina L. under strictly non-inductive photoperiods and hasten it under inductive photoperiods, cyclic AMP being more effective than 3- or 5-AMPs in this regard.Abbreviations cyclic AMP cyclic 3,5-adenosine monophosphate  相似文献   

12.
A protein kinase that phosphorylates histones and polysomal proteins was partially purified from mouse liver cytosol. The active enzyme has a molecular mass of 100 kDa and a phosphorylatable subunit of 54 kDa. Biochemical as well as immunological data suggest that the enzyme is a heterodimer composed of the catalytic subunit of cyclic AMP-dependent protein kinase and the RII regulatory subunit. This RC form does not seem to dissociate upon activation with 3, 5 cyclic AMP and exhibits identical specificity as the classical cAMP-dependent protein kinase (2.7.1.37). The enzyme is affected by the 3, 5 cyclic phosphates of adenosine mainly, but also of guanosine, uridine and cytidine in a substrate-dependent manner. Cyclic nucleotides slightly stimulate phosphate incorporation into histones, while phosphorylation of polysomal proteins in intact polysomes is dramatically increased. The substrate- specific stimulatory effects of 3, 5 cyclic nucleotides are due to repression of the inhibition exerted upon the reaction, by negatively charged macromolecules such as RNA, DNA and to a lesser extent heparin.  相似文献   

13.
Summary Clear-plaque phage c, attacking bacitracin-producing strains of B. licheniformis, yields spontaneous temperate mutants at high frequency; the temperate mutants fall into several classes phenotypically different in plaque morphology and properties of lysogenised bacteria. The most common phenotype 3 has DNA restriction fragment patterns identical with those of the parent c; some less common temperate forms, i.e. 1 and 2, produce different restriction fragment patterns, sugesting that a part of the original c DNA has been reorganized or replaced by some foreign genetic material. The changed fragment pattern remains stable upon subsequent passaging of the phage or of the lysogenic bacteria. Neither class of temperate phage mutants gives clearplaque revertants at measurable frequency. Lysogenisation of bacteria with any class of temperate phage confers immunity to all temperate forms and to c; virulent mutants vir, which plate with 100% efficiency on lysogens for 1 and 2 but not for 3, occur in stocks of c at a frequency of 10–7. The mutation from c to vir is not accompanied by any change of the restriction fragment patterns of DNA.  相似文献   

14.
Pilobolus longipes spores were activated by either glucose or 6-deoxyglucose. Glucose-induced spore activation was previously shown to follow an increase in intracellular cyclic AMP. Concurrent with glucose-induced spore activation, were shifts in 6-deoxyglucose transport kinetics towards higher V max and K m values. Cyclic AMP derivatives also caused spore activation and similar changes in the kinetic parameters of 6-deoxyglucose transport. The time course of activation was paralleled by changes in transport activity. Inhibition of phosphodiesterase alone did not cause activation or induce changes in transport activity, but in combination with sub-optimal levels of either 6-deoxyglucose or cAMP derivatives, it amplified the germination signals to produce large increases in both spore activation and 6-deoxyglucose transport activity. These results support the conclusion that glucose transport in germinating spores is regulated by cAMP.Abbreviations IBMX 3-isobutyl-1-methylxanthine; monobutyryl cyclic AMP - N6 monobutyryladenosine 3:5-cyclic monophosphate - 8-bromo cyclic AMP 8-bromoadenosine 3:5-cyclic monophosphate  相似文献   

15.
The relative contribution of autotrophic carbon sources (aquatic macrophytes, flooded forest, phytoplankton) for heterotrophic bacterioplankton was evaluated in a floodplain lake of the Central Amazon. Stable carbon isotopes (13C) were used as tracers. Values of 13C of different autotrophic sources were compared to those of dissolved organic carbon (DOC) and those of bacterially produced CO2.The percentage of carbon derived from C4 macrophytes for bacterially produced CO2 was the highest, on average 89%. The average 13C value of CO2 from bacterial respiration was –18.5 ± 3.3. Considering a fractionation of CO2 of 3 by bacterial respiration, 13C value was –15.5, near C4 macrophyte 13C value (–13.1).The average value of total DOC 13C was –26.8 ± 2.4. The percentage of C4 macrophytes carbon for total DOC was on average 17%. Considering that bacteria consume mainly carbon from macrophytes, the dominance of C3 plants for total DOC probably reflects a faster consumption of the former source, rather than a major contribution of the latter source.Heterotrophic bacterioplankton in the floodplain may be an important link in the aquatic food web, transferring the carbon from C4 macrophytes to the consumers.  相似文献   

16.
The acoustic near field of a dancing honeybee   总被引:1,自引:0,他引:1  
Summary The acoustic near field close to honeybees performing the wagging dance was investigated with pairs of small, matched microphones placed in various positions around the dancing bees. The dance sounds are produced by the wings, which act as an asymmetrical dipole emitter. Close to the abdomen, the sound pressures in the air spaces above and below the plane of the wings are totally out of phase. A zone of very intense acoustical short-circuiting exists close to the edges of the wings, where pressure gradients of about 1 Pa/mm are observed in the dorso-ventral direction (perpendicular to the plane of the wings). The pressure gradients drive air movements with velocity amplitudes up to about 1 m/s. The pressure gradients are much smaller in directions radially away from the bee and decrease rapidly with increasing distance from the wings. The sound pressure detected by a stationary probe at one side of the bee is strongly modulated at 12–13 Hz as a result of the bee's side-to-side wagging. Surprisingly little sound is found near the dancer's head. The positions of the follower bees reflect the properties of the acoustic field: The follower bees place their antennae in the zone of maximum acoustical short-circuiting where the air particle movements are most intense. These observations suggest 1) how follower bees can avoid mixing up the messages carried by the dance sounds when two or more bees are dancing only a few cm apart and 2) how the followers might extract information about a dancer's spatial orientation from the acoustic near field she produces. The observations also provide clues regarding the nature of the putative sound receivers.Dedicated to Professor Dr. Drs. h.c. mult. H. Autrum on the occasion of his 80th birthday  相似文献   

17.
A human gene previously identified as a partial cDNA homologous to the gene of RET finger protein was characterized. Northern hybridization detected three messages of 3.3, 4.2, and 7.5kb. The coding sequences of the more abundant of the three messages, the 4.2 and the 3.3kb, were determined. The former encodes a 630 amino acid protein (TRIM41) and the latter a 518 amino acid protein (TRIM41). Green fluorescent protein (GFP) fusions of full-length TRIM41 and TRIM41 were both observed as speckles in the cytoplasm and the nucleus. The result was corroborated by Western analysis of cellular fractions. Results with GFP fusions of various segments of the TRIM41 proteins indicated that the nuclear transport of the proteins is mediated by an N-terminal segment common to both isoforms, but independent of a classical nuclear localization signal sequence.  相似文献   

18.
Phosphomonoesterase and diesterase that cleave phosphatidylinositol-4-phosphate (diphosphoinositide, DPI) and phosphatidylinositol-4,5-bisphosphate (triphosphoinositide, TPI) were detected in three subfractions of purified rat brain myelin, and some properties of the enzymes were studied. Monoesterase activity was stimulated by KCl, maximally at a concentration of 25 mM, and inhibited at KCl concentrations above 50 mM. Addition of boiled pH 5 supernatant of rat brain homogenate doubled the enzymic activity; EDTA was inhibitory. The specific activities were nearly equal in the low density, medium density, and heavy density myelin fractions but about 30% lower than in whole brain homogenate. The monophosphatase could be solubilized by extraction with 0.2% Triton X-100. The phosphodiesterase activity was inhibited by EDTA and EGTA and not stimulated by KCl or pH 5 supernatant. Specific activities were nearly equal in whole brain and myelin but were by about 60 percent elevated in the heavy density over the low density myelin fraction. These results show that the hydrolases operative in the fast turnover of the inositide phosphate groups are distributed over the entire myelin structure.  相似文献   

19.
Numerous barley cultivars from around the world have been identified as potential sources of Fusarium head blight (FHB) resistance genes. All of these cultivars exhibit partial resistance, and several mapping studies have shown that resistance to FHB is controlled by multiple genes. Successful development of barley cultivars with high levels of FHB resistance will require combining genes from multiple sources. We characterized five potential new sources of FHB resistance (AC Oxbow, Atahualpa, HOR211, PFC88209, and Zhedar#1) to determine if they contain new FHB resistance genes. Cluster analysis, using a set of 80 SSR markers distributed throughout the genome, showed that most of the new sources of resistance were not similar to three cultivars that have been used in previous FHB mapping studies (Chevron, Frederickson, and Gobernadora), with Atahualpa and HOR211 being the most dissimilar. By selective genotyping, we determined whether markers linked to six known FHB resistance quantitative trait loci (QTLs), discovered in other genotypes, explained variation for resistance in advanced breeding populations created from the new sources of resistance. Markers linked to four of the six known QTLs were associated with FHB severity in at least one of the populations. However, none of the six QTL regions were associated with variation for FHB severity in populations derived from crosses that utilized sources of resistance HOR211 or PFC88209. Selective genotyping is an efficient method for breeders to utilize current QTL information about disease resistance to search for new resistance genes.  相似文献   

20.
Purified ribulose-bisphosphate carboxylase (EC 4.1.1.39) was strongly and equally inhibited either by ADP or GDP and to a lesser extent by IDP. AMP or ATP exerted little effect on activity. Inhibition by the nucleotide diphosphates was competitive with respect to RuBP and non-competitive with respect to CO2 and Mg2+, respectively. Treatment of the enzyme with urea or guanidine-HCl resulted in rapid loss of activity that was not restored by dialysis even in the presence of Mg2+ and cysteine. Sodium dodecyl sulfate electrophoresis of 8.0 M urea treated enzyme revealed the presence of a fast-moving (small) sub-unit with molecular weight 14150 and a slower moving (large) sub-unit with molecular weight 68000. Examination of native enzyme by sodium dodecyl sulfate electrophoresis gave sub-units of 13700 and 55500 respectively. The amino acid content standardized to phenylalanine was essentially similar to that from other sources. Arrhenius plots showed a break at 29°C with an E a of 12.34 kcal per mole for the steeper part of the curve and a H of 11.43 kcal per mole while for the less steep region, the E a was 1.04 kcal per mole and the H 1.92 kcal per mole.Abbreviations ADP adenosine-5-diphosphate - AMP adenosine-5-monophosphate - ATP adenosine-5-triphosphate - CDP cytidine-5-diphosphate - CMP cytidine-5-monophosphate - CTP cytidine-5-triphosphate - FDP fructose-1,6-diphosphate - F6P fructose-6-phosphate - GDP guanosine-5-diphosphate - GMP guanosine-5-monophosphate - G6P glucose-6-phosphate - GTP guanosine-5-triphosphate - IDP inosine-5-diphosphate - IMP inosine-5-monophosphate - PEP phosphoenolpyruvate - 6PG 6-phosphogluconate - R1P ribose-1-phosphate - R5P ribose-5-phosphate - RuBP ribulose-1,5-bisphosphate - SDS sodium dodecyl sulfate - TDP thymidine-5-diphosphate - TMP thymidine-5-monophosphate - TTP thymidine-5-triphosphate - UDP uridine-5-diphosphate - UMP uridine-5-monophosphate - UTP uridine-5-triphosphate  相似文献   

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