• Title, Summary, Keyword: AMP

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The effect of cyclic AMP on the growth of Toxoplasma gondii in vitro (Cyclic AMP대사가 Toxoplasma gondii의 체외 배양에 미치는 영향)

  • 최원영;남호우
    • The Korean Journal of Parasitology
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    • v.28 no.2
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    • pp.71-78
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    • 1990
  • To assess the role of cAMP on the growth and proliferation of Toxoplasma in HL-60 cells we tested the effect of exogenous cAMP and cAMP analogues to the co-culture system of Toxoplasma and HL-60 cells. cAMP, dbcAMP, and br-cAMP stimulated the growth of Texoplasma at a specific concentration, i.e., 100 mM, l00 mM, and 10-1 mM, respectively. There were differences in growth induction kinetics and in the rate of promotion. These results were further verified by treating the co-culture with adenylate cyclase activator, pNHppG, cAMP phosphodiesterase activators, imidasole and A23187, and cAMP phosphodiesterase inhibitors, IBMX, compound 48/80, and theophylline, separately. When the cytosolic cAMP levels increased by the reagents mentioned above, Toxoplasma in the cytoplasm of HL-60 cells stimulated to proliferate more rapidly with concentration-dependent modes compared to the control, and vice versa. It is suggested that some mechanisms are activated by the high levels of cAMP in the cytoplasm, which result in the stimulation of Toxoplasma proliferation.

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Cyclic AMP Receptor Protein Adopts the Highly Stable Conformation at Millimolar cAMP Concentration (높은 cAMP 농도에서 cAMP 수용성 단백질의 열 안정화)

  • Kang, Jong-Baek;Choi, Young
    • Journal of Life Science
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    • v.13 no.5
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    • pp.751-755
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    • 2003
  • Cyclic AMP receptor proteins(CRP) activate many genes in Escherichia coli by binding of cAMP with not fully known mechanism. CRP existed as apo-CRP in the absence of cAMP, $CRP;(cAMP)_2$$_2$ at low(micromolar) cAMP concentration, or $CRP;(cAMP)_4$ at high(millimolar) concentration of cAMP. This study is designed to measure the thermal stability of S83G CRP, which substituted glycine for serine at amino acid 83 position, with CD spectrapolarimeter at 222nm by the constant elevation of temperature from $20^{\circ]C\; to\; 90^{\circ}C\; at\; 1^{\circ}C/min$. The non-linear regression analysis showed that melting temperatures were 68.4, 72.0, and $82.3^{\circ}C$ for no cAMP, 0.1mM cAMP, and 5mM cAMP, respectively. Result showed the strong thermal stability of CRP by binding of additional cAMP molecules to region between the hinge region and helix-turn-helix(HTH) motif at 5mM cAMP concentration.

Studies on the Effects of cAMP on the ATPase Activity and on the Calcium Uptake of the Sarcoplasmic Reticulum (근 소포체의 ATPase 활성과 Ca 능동수송에 미치는 cAMP의 영향)

  • 河斗鳳;朴姬淳;尹炳宇;金漢都
    • The Korean Journal of Zoology
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    • v.18 no.4
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    • pp.221-229
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    • 1975
  • The effect of adenosine 3', 5'-cyclic monophosphate on the ATPase activity and on the active transport of Ca of the sarcoplasmic reticulum fragments of the rabbit skeletal muscle was studied. Cyclic AMP (cAMP) had no effect on the ATPase activity of the fragments (8,000 ~ 20,000 $\times$ G and 20,000 ~ 36,000 $\times$ G fractions). $N^6$, O^{2'} -Dibutyryl cAMP (DBcAMP) had either no effect on the activity. On the other hand, theophylline (1 mM) increased the activity by about 20%. The active uptake of Ca by the sarcoplasmic reticulum fragments was inhibited by the presence of 1$\times$$10^{-6}$ ~ 1 $\times$ $10^{-3}$M of cAMP. The presence of DBcAMP or theophylline also inhibited the uptake. It is, therefore, concluded that the Ca uptake of the sarcoplasmic reticulum seems to be controlled by cAMP.

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Elliptic Curve AMP Protocol (타원곡선을 이용한 AMP 프로토콜)

  • Ahn, Chang-Sup;Heu, Shin
    • Journal of KIISE:Computer Systems and Theory
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    • v.29 no.11
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    • pp.622-633
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    • 2002
  • Authentication and Key Agreement using password provide convenience and amenity, but what human can remember has extremely low entropy. To overcome its defects, AMP(Authentiration and key agreement via Memorable Password) which performs authentication and key agreement securely via low entropy password are presented. AMP uses Diffie-Hellman problem that depends on discrete logarithm problem. Otherwise, this thesis applies elliptic curve cryptosystem to AMP for further efficiency That is, this thesis presents EC-AMP(Elliptic Curve-AMP) protocol based on elliptic curve discrete logarithm problem instead of discrete logarithm problem, and shows its high performance through the implementation. EC-AMP secures against various attacks in the random oracle model just as AMP Thus, we nay supply EC-AMP to the network environment that requires authentication and key agreement to get both convenience and security from elliptic curve discrete logarithm problem.

Uptake and Degradadon of cAMP by Frog Follides in vitro (배양중인 개구리 여포의 cAMP 흡수와 분해)

  • 권혁방;나철호;안련섭;김경진
    • The Korean Journal of Zoology
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    • v.34 no.2
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    • pp.181-187
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    • 1991
  • We previously showed that transient exposure of Rana dybowskii follicles to exogenous cAMP in vitro could induce meiotic maturation. The present experiments were carried out to acertain whether the exogenous cAMP penetrate into the follicles. Isolated follicles were precultured in the medium containing cAMP (2.5 mM) for 6 hours and then cultured further in plain medium for 18 hours. The change of intrafollicular cAMP levels during the culture period were examined by utilizing cAMP radioimmunoassay (RIA). The intrafollicular levels of cAMP increased about thirty times of the basal level (about 3 p mole/follicle) in two hours and reached a peak in six hours (170 p mole/follicle) during the preculture period. However, when the follicles were transferred to plain medium, the levels decreased markedly in six hours to very low levels (about 10 p mole/ follicle), and kept the same levels thereafter. But the levels did not decrease to the basal levels. The increase and decrease of the intrafollicular cAMP was not affected by the presence of isobutyl methylxanthine (IBMX) or progesterone. The data suggest that exogenous cAMP pene-trate into the follicles and the cAMP accumulated by the follicles are degraded very rapidly.

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Involvement of the Cyclic AMP-Protein Kinase A Pathway in Gap Junctional Communication in Preimplantation Mouse Embryos

  • Haengseok Song;Gye, Myung-Chan;Jun, Jin-Hyun
    • Animal cells and systems
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    • v.2 no.1
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    • pp.99-106
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    • 1998
  • In this study, we have examined the role of cAMP in gap junctional communication (GJC) in preimplantation mouse embryos. GJC was monitored by Lucifer Yellow (LY) injected into one blastomere of compacted embryos. The speed of GJC was defined as the time taken for the last blastomere of the embryo to become visibly fluorescent. The median time for 8-cell embrvos (140 sec) was similar to that for 16-cell (135 sec). To determine whether cAMP and cAMP-dependent protein kinase (PKA) are involved in the regulation of GJC, the effects of PKA inhibitor (H8) and cAMP analogues (Rp-cAMP and 8-Br-cAMP) on dye transfer between blastomeres of compacted embryos were examined. Some of the embryos treated with either H8 or Rp-cAMP failed to transfer LY to all blastomeres within 10 min. In contrast, 8-Br-cAMP speeded up fluorescent dye transfer. The median time to fill all blastomeres with LY was 140 sec in untreated controls and 90 sec in siblings treated with 8-Br-cAMP. Inhibition of PKA by H8 or Rp-cAMP induced delay or arrest in embryo development after compaction, but the increase of intracellular cAMP showed no effect. These findings suggest that GJC in preimplantation mouse embryos is regulated by cAMP-PKA pathway and transient interference by PKA inhibitors induces the developmental delay beyond compaction.

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Immunomodulatory Effect of cAMP-Elevating Agents on Macrophage- and T cell-Mediated Immune Responses (cAMP 증가 유도 약물의 대식세포- 및 T 세포-매개성 면역반응 조절작용)

  • Rhee, Man-Hee;Cho, Jae-Youl
    • YAKHAK HOEJI
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    • v.51 no.1
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    • pp.35-43
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    • 2007
  • To investigate the immunomodulatory roles of cyclic AMP (CAMP) on macrophage- and T lymphocyte-mediated immune responses, CAMP elevating agents were employed and carefully re-examined under the activation conditions of the cells. Various inhibitors tested dose-dependently blocked tumor necrosis factor (TNF)-${\alpha}$ production with IC$_{50}$ values ranged from 0.04 to 300 ${\mu}$M. Of the inhibitors, cAMP-elevating agents showed lower cytotoxicity assessed by lactate dehydrogenase (LDH) release, suggesting less toxic and more selective. In particular co-treatment of dbcAMP with a protein kinase C inhibitor staurosporine displayed the synergistic inhibition of TNF-${\alpha}$ production. The modulatory effect of dbcAMP on TNF-${\alpha}$ and nitric oxide (NO) was significantly affected by treatment time of dbcAMP. Thus, post-treatment of dbcAMP (three hours before LPS) abrogated dbcAMP's inhibitory activity and rather enhanced TNF-${\alpha}$ level up to 60%. In contrast, additional NO production was shown at the co-treatment of dbcAMP with LPS. Unlike simultaneous treatment of phorbol 12-myristate 13-acetate (PMA) and interferon (IFN)-${\gamma}$co-treatment, the combination of dbcAMP with other NO-inducing stimuli did not show drastic overproduction of NO. cAMP elevating agents also diminished splenocyte proliferation stimulated by concanavalin (Con) A, phytohemaglutinin A (PHA) and lipopolysaccharide (LPS). In addition, dbcAMP but not rolipram strongly suppressed CD8$^+$ T cells (CTLL-2). Finally, cAMP elevating agents were differentially involved in regulating CD98-mediated cell-cell adhesion. Thus, dbcAMP and rolipram significantly enhanced the cell-cell adhesion, whereas forskolin blocked. Therefore, our results suggest that CAMP elevating agents participate in various immune responses mediated by macrophages and T cells with a different fashion depending on cellular environments and activation signals.

Studies on the Effects of Dibutyryl Cyclic AMP and Theophylline on RNA Synthesis in Mouse Follicular Oocytes in Vitro (Dibutyryl Cyclic AMP와 Theophylline이 培養중인 생쥐 濾胞卵子의 RNA合成에 미치는 영향에 관한 연구)

  • Cho, Wan-Kyoo;Yoo, Seung-Hyun
    • The Korean Journal of Zoology
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    • v.18 no.1
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    • pp.19-26
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    • 1975
  • 自記放射法을 이용하여 dbcAMP와 theophylline이 未成熟卵子의 RNA合成에 미치는 영향을 관찰하였다. 培養中인 未成熟卵子의 RNA合成은 dbcAMP와 theophylline에 의하여 抑制를 받았다. dbcAMP나 theophylline은 培養液(modified Krebs-Ringer bicarbonate solution)內에 100 $\\mu$g/ml 정도 들어 있으며 핵막(germinal vesicle)의 붕괴 되지 못하고 그대로 存在하며 그동안의 RNA合成은 극히 억제된 채로 남아 있다. 그러나 培養을 시작하여 2$\\sim$3時間後, 즉 핵막붕괴가 끝난 다음에 이들 억제물질을 배양액에 添加하면 正常卵子와 같이 성숙분열이나 RNA合成이 억제 됨이 없이 진행된다. 24時間동안 dbcAMP나 theophylline으로 성숙이 억제 되었던 卵子도 이들 억제물질을 제거하면 즉시 成熟分裂이 진행되며 RNA合成도 正常的으로 일어난다. 이런 結果로 미루어 dbcAMP등의 RNA合成 抑制機作에 한 가지 가능성을 추측할 수 있다. 즉 dbcAMP나 theophylline의 처리에 의해 細胞質內 cAMP의 농도가 높아지고 이 cAMP는 핵막붕괴나 染色質의 응집에 관여하는 단백질 合成을 誘導할 mRNA合成을 억제하며 이 때문에 卵子는 핵을 보유한채 그대로 남아 있는 것이다.

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Vasodilating Mechanism of Dibutyryl-cAMP and Forskolin in Rabbit Aorta (Dibutyryl-cyclic AMP와 Forskolin의 혈관평활근 이완작용)

  • Ahn, Hee-Yul;Lim, Jung-Kyoo
    • The Korean Journal of Pharmacology
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    • v.26 no.2
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    • pp.127-133
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    • 1990
  • Dibutyryl-cyclic AMP (db-cAMP) and forskolin were used to investigate vasodilating mechanism of cAMP in rabbit aorta. Db-cAMP and forskolin inhibited the development of contractile tension induced by norepinephrine (NE) concentration-dependently. However, high $K{^+}-induced$ contractile tension was inhibited less effectively by db-cAMP and forskolin. Db-cAMP and forskolin inhibited $^{45}Ca^{2+}$ uptake increased by NE. Forskolin seemed to inhibit $^{45}Ca^{2+}$ uptake increased by high $K{^+}$, but this inhibition was not significant statistically. Db-cAMP inhibited $Ca^{2+}-transient$ contraction by NE in $Ca^{2+}-free$ solution. In conclusion, it seems that cAMP blocks $Ca^{2+}$ influx through receptor operated $Ca^{2+}$ channels (ROCs), but that the effect of cAMP on $Ca^{2+}$ influx through voltage gated $Ca^{2+}$ channels (VGCs) is not clear in this experiment. Furthermore, cAMP is likely to inhibit calcium release from the intracellular stores.

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Functional Characterization of cAMP-Regulated Gene, CAR1, in Cryptococcus neoformans

  • Jung, Kwang-Woo;Maeng, Shin-Ae;Bahn, Yong-Sun
    • Mycobiology
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    • v.38 no.1
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    • pp.26-32
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    • 2010
  • The cyclic AMP (cAMP) pathway plays a major role in growth, sexual differentiation, and virulence factor synthesis of pathogenic fungi. In Cryptococcus neoformans, perturbation of the cAMP pathway, such as a deletion in the gene encoding adenylyl cyclase (CAC1), causes defects in the production of virulence factors, including capsule and melanin production, as well as mating. Previously, we performed a comparative transcriptome analysis of the Ras- and cAMP- pathway mutants, which revealed 163 potential cAMP-regulated genes (38 genes at a 2-fold cutoff). The present study characterized the role of one of the cAMP pathway-dependent genes (serotype A identification number CNAG_ 06576.2). The expression patterns were confirmed by Northern blot analysis and the gene was designated cAMP-regulated gene 1 (CAR1). Interestingly, deletion of CAR1 did not affect biosynthesis of any virulence factors and the mating process, unlike the cAMP-signaling deficient cac1$\Delta$ mutant. Furthermore, the car1$\Delta$ mutant exhibited wild-type levels of the stress-response phenotype against diverse environmental cues, indicating that Car1, albeit regulated by the cAMP-pathway, is not essential to confer a cAMP-dependent phenotype in C. neoformans.