• Title, Summary, Keyword: ATP

Search Result 1,653, Processing Time 0.061 seconds

Immobilization of ATP on Bovine $\beta$- Caseins by Using Transglutaminase (효소법에 의한 ATP의 Bovine $\beta$-Casein에의 고정화)

  • 윤세억;박선영김명곤
    • KSBB Journal
    • /
    • v.5 no.3
    • /
    • pp.241-246
    • /
    • 1990
  • ATP analogs were immobilized or bovine caseins by the action of transglutaminase. The ATP analogs immobilized on the caseins were enzymatically active and interconverten by kinases. The immobilized ATP was dephosphorylated to the corresponding ADP by hexokinase and rephosphorylated to the ATP in solid form by acetate kinase. Under the conditions chosen, about 55% of the immobilized ATP was dephosphorylated and about 80% of the resulted ADP was rephosphorylated. Bovine $\beta$-casein was more useful than $\alpha$sf-casein as a carrier and C8-substituted ATP analognwas more effective than N6-substituted one in immobilization. Michaelis constant of C8-substituted ATP analog immobilized on $\beta$-casein was similar to that of free form of ATP and that of ATP analog. The immobilized ATP was much more stable than free ATP and its analog, while maximum velocity was reduced to 37% of the free ATP analog. The immobilized ATP was recovered almost completely by calcium precipitation.

  • PDF

Distribution of ATP in the Deep-Sea Sediment in the KODOS 97-2 Area, Northeast Equatorial Pacific Ocean (북동적도 태평양 KODOS 97-2 해역 심해저 퇴적물 내의 ATP 분포양상)

  • Hyun, Jung-Ho;Kim, Kyeong-Hong;Chi, Sang-Bum;Moon, Jai-Woon
    • The Sea
    • /
    • v.3 no.3
    • /
    • pp.142-148
    • /
    • 1998
  • Environmental baseline information is necessary in order to assess the potential environmental impact of future manganese-nodule mining on the deep-seabed ecosystem. Total ATP (T-ATP), dissolved ATP (D-ATP) and particulate ATP (P-ATP) were measured to estimate total microbial biomass and to elucidate their vertical distribution patterns in the seabed of KODOS (Korea Deep Ocean Study) area, northeast equatorial Pacific Ocean. Within the upper 6 cm depth of sediment, the concentrations of T-ATP, D-ATP and P-ATP ranged from 4.4 to 40.6, from 0.6 to 16.1, and from 3.0 to 29.2 ng/g dry sediment, respectively. Approximately 84% of T-ATP, 81% of D-ATP, and 74% of P-ATP were present within the topmost 2 cm depth of sediment, and the distributions of ATP were well correlated with water content in the sediment. These results indicate that the distribution of total microbial biomass was largely determined by the supply of organic matter from surface water column. Fine-scale vertical variations of ATP were detected within 1-cm thick veneer of the sediment samples collected by multiple corer, while no apparent vertical changes were observed in the box-cored samples. It is evident that the box-core samples were disturbed extensively during sampling, which suggests that the multiple corer is a more appropriate sampling gear for measuring fine-scale vertical distribution pattern of ATP within thin sediment veneer. Overall results suggest that the concentrations of ATP, given their clear changes in vertical distribution pattern within 6 cm depth of sediment, are a suitable environmental baseline parameter in evaluating the variations of benthic microbial biomass that are likely to be caused by deep-seabed mining operation.

  • PDF

ATP and Purinergic Receptor Agonists Stimulate the Mitogen-Activated Protein Kinase Pathway and DNA Synthesis in Mouse Mammary Epithelial Cells

  • Yuh In-Sub
    • Reproductive and Developmental Biology
    • /
    • v.28 no.4
    • /
    • pp.211-219
    • /
    • 2004
  • The effects of adenosine 5'-triphosphate (ATP) and ATP analogs, P/sub 2y/ purinoceptor agonists, on growth of normal mouse mammary epithelial cells (NMuMG) were examined. Cells were plated onto 24 well plates in DMEM supplemented with 10 % fetal calf serum. After serum starvation for 24 hours, ATP, P/sub 2y/ purinoceptor agonists (AdoPP[NH]P, ATP-α-S, ATP-γ-S, β, γ-me-ATP and 2me-S-ATP), P/sub 2u/ purinoceptor agonist (UTP) and P/sub 2y/ purinoceptor antagonists (Reactive Blue 2, more selective to P/sub 2y/ receptor than PPADS; PPADS) were added. DNA synthesis was estimated as incorporation of 3H-thymidine into DNA (1 hour pulse with 1 μ Ci/ml, 18~19 hours after treatment). ATP, Adopp[NH]P, ATP-α-S or ATP-γ-S, significantly increased DNA synthesis at 1, 10 and 100 μM concentrations with dose-dependency (P<0.05), and the maximum responses of ATP and ATP analogs were shown at 100 μM concentration (P<0.05). The potency order of DNA synthesis was ATP≥ATP- γ -S>Adopp [NH]P>ATP-α-S. β, γ -me-ATP, 2me-S-ATP and UTP did not increase DNA synthesis. In autoradiographic analysis of percentage of S-phase cells, similar results were observed to those of DNA synthesis. Addition of 1, 10 or 100 μM Reactive Blue 2 or PPADS significantly decreased ATP (100 μM)-induced DNA synthesis, however, PPADS was less effective than Reactive Blue 2. In Elvax 40P implant experiment, ATP directly stimulated mammary endbud growth in situ suggesting the physiological regulator of ATP in mammary growth. ATP 100 μM rapidly increased MAPK activity, reaching a maximum at 5 min and then gradually decreasing to the base level in 30 min. ATP analogs, Adopp[NH]P and ATP-γ-S also increased MAPK activity, however, β, γ-me-ATP and 2me-S-ATP did not. The inhibitor of the upstream MAPK kinase (MEK), PD 98059 (25 μM), effectively reduced ATP (100 μM) or EGF(10 ng/ml, as positive control)-induced MAPK activity and DNA synthesis (P<0.05). These results indicate that ATP-induced DNA synthesis was prevented from the direct inhibition of MAPK kinase pathway. Overall results support the hypothesis that the stimulatory effects of normal mouse mammary epithelial growth by addition of ATP or ATP analogs are mediated through mammary tissue specific P/sub 2y/ purinoceptor subtype, and MAPK activation is necessary for the ATP-induced cell growth.

Development of ATP Train Separation Control Simulator for Radio-based Train Control System (무선통신기반 열차제어시스템 ATP 열차간격제어알고리즘 시뮬레이터 개발)

  • Yoon, Yong-Ki;Oh, She-Chan;Choi, June-Young;Park, Jae-Young;Yang, Hai-Won
    • Journal of the Korean Society for Railway
    • /
    • v.15 no.1
    • /
    • pp.29-36
    • /
    • 2012
  • This paper includes ATP(Automatic Train Protection) simulator development and ATP algorithm verification to allocate wayside and train-borne and verify ATP functions of communications based train control system. The train control system has some characteristics such as simple structure and high safety when wireless communication technology is applied to the train control system. Especially, vital functions can be performed with in wayside and train-borne ATP. However, different system can be realized because I/F contents vary in accordance with vital functional allocation of ATP. Drawing characteristics in accordance with wayside and train-borne functional allocation and drawing I/F details affected by such characteristics are needed accordingly. This paper includes ATP simulator development creating train location information by direct activation of an electric motor, verifies train safety distance control algorithm of ATP by functional allocation such as train movement authority and train speed limit to ATP, and draws any supplementation needed. Appropriate simulated environment for verify ATP algorithm and main factors that affect to the ATP function were confirmed.

Intragenic DNA Methylation Concomitant with Repression of ATP4B and ATP4A Gene Expression in Gastric Cancer is a Potential Serum Biomarker

  • Raja, Uthandaraman Mahalinga;Gopal, Gopisetty;Rajkumar, Thangarajan
    • Asian Pacific Journal of Cancer Prevention
    • /
    • v.13 no.11
    • /
    • pp.5563-5568
    • /
    • 2012
  • Based on our previous report on gastric cancer which documented ATP4A and ATP4B mRNA down-regulation in gastric tumors relative to normal gastric tissues, we hypothesized that epigenetic mechanisms could be responsible. ATP4A and ATP4B mRNA expression in gastric cancer cell lines AGS, SNU638 and NUGC-3 was examined using reverse transcriptase PCR (RT-PCR). AGS cells were treated with TSA or 5'-AzaDC and methylation specific PCR (MSP) and bisulfite sequencing PCR (BSP) analysis were performed. MSP analysis was on DNA from paraffin embedded tissues sections and plasma. Expression analysis revealed downregulation of ATP4A and ATP4B genes in gastric cancer cell lines relative to normal gastric tissue, while treatment with 5'-AzaDC re-activated expression of both. Search for CpG islands in their putative promoter regions did not indicate CpG islands (CGI) but only further downstream in the bodies of the genes. Methylation specific PCR (MSP) in the exon1 of the ATP4B gene and exon7 in ATP4A indicated methylation in all the gastric cancer cell lines tested. MSP analysis in tumor tissue samples revealed methylation in the majority of tumor samples, 15/19, for ATP4B and 8/8 for ATP4A. There was concordance between ATP4B and ATP4A down-regulation and methylation status in the tumour samples tested. ATP4B methylation was detectable in cell free DNA from gastric cancer patient's plasma samples. Thus ATP4A and ATP4B down-regulation involves DNA methylation and methylated ATP4B DNA in plasma is a potential biomarker for gastric cancer.

Alteration of Biochemical Responses in Activated Human Neutrophils by ATP and Adenosine (활성화된 사람 중성 백혈구에서 ATP와 Adenosine 처리에 따른 생화학적 반응의 변경)

  • Park, Sung-Soo;Lee, Chung-Soo
    • The Korean Journal of Pharmacology
    • /
    • v.26 no.1
    • /
    • pp.55-66
    • /
    • 1990
  • In both resting and opsonized zymosan activated neutrophils, ATP stimulated superoxide generation, whereas adenosine inhibited it slightly. The superoxide generation in activated neutrophils to ATP was greater than that of resting neutrophils. In $Ca^{++}$ free medium, inhibitory effect of adenosine on superoxide generation was detectable, whereas ATP did not have any effect. The stimulatory effect of ATP on superoxide generation was inhibited by adenosine in a dose dependent manner. Neither ATP nor adenosine had any effect on NADPH oxidase acitivity. Effects of ATP or adenosine on superoxide generation were more prominent than that by other triphosphate nucleotides or nucleosides. ATP and ADP further stimulated $Ca^{++}$ uptake and increased cytosolic free $Ca^{++}$ level in neutrophils activated by opsonized zymosan, but adenosine inhibited a $Ca^{++}$ mobilization. Verapamil effectively and tetrodotoxin slightly inhibited an increase of cytosolic free $Ca^{++}$ level induced by ATP. Inhibitory effect of either verapamil or tetrodotoxin on superoxide generation in the ATP plus opsonized zymosan-activated neutrophils was greater than in the cells activated by opsonized zymosan alone. Tetraethylammonium chloride had no apparent effect on superoxide generation. CCCP, 2,4-dinitrophenol, diphenylhydantoin and procaine all inhibited superoxide generation in neutrophils activated by opsonized zymosan. Among these, CCCP only inhibited a stimulatory effect of ATP. ATP further stimulated a loss of sulfhydryl groups in activated neutrophils, whereas adenosine had no effect on it. These results suggest that functional responses of neutrophils may be regulated at least partly by purines. ATP and adenosine may further after functional responses of activated neutrophils through their effect on $Ca^{++}$ uptake, membrane phosphorylation and oxidation of soluble sulfhydryl groups.

  • PDF

Development of Microfluidic Channel for Pretreatment of Extracellular ATP using DEP Force (DEP를 이용한 세포 외부 ATP 제거 전처리 미세 유로의 개발)

  • Lim, Hee-Taek;Jung, Hyo-Il
    • Proceedings of the KSME Conference
    • /
    • /
    • pp.1687-1689
    • /
    • 2008
  • In the detection of pathogenic microorganisms ATP-bioluminescence reaction is a fascinating method. ATP(adenosine triphosphate) is an energy source of all kinds of living organism and ATP-bioluminescence reaction uses this ATP. However, ATP exists not only in the cells but also outside the cells. Therefore ATP-bioluminescence reaction only with intracellular ATP is very important in pathogenic microorganism detection. Because of that reason we developed a microfluidic channel containing Dielectrophoretic zone which capture microorganisms and eliminating and washing extracellular ATP with ATP-degarading enzymes, adenosine phosphate deaminase and apyrase. Microorganisms are captured by pDEP force at the DEP electrode zone and only extracellular ATPs are washed and eliminated outside the zone.

  • PDF

ATP Model Related CRM in SCM Environment (SCM환경에서 CRM을 이용한 ATP 모델 연구)

  • 박주식;김원식;남호기;박상민
    • Journal of the Korea Safety Management and Science
    • /
    • v.3 no.1
    • /
    • pp.45-56
    • /
    • 2001
  • In the supply chain, The ATP function doesn't only give customers to confirmation of delivery. It can be used by the core function with ATP rule that can reconcile supplies and demands on the supply chain. Therefore We can acquire the conformation about accuracy on the due date of supplier by using the ATP function of management about real and concurrent access on the supply chain, also can decide the affect about product availability due to forecasting or customer's orders through the ATP. This study analyze the data concerned with ATP and define the necessity on a SCM solution. Under the these environments, after defining the ATP rule that can improve the customer value and data flow related the CRM, we propose the advanced ATP model that proposes the method and classification system that can flexibly aggregate the ATP data with ATP rule on the supply chain.

  • PDF

Synthesis and Photoaffinity Labeling of 3'(2')-O-(p-azidobenzoyl) ATP

  • Shin, Seung-Jin;Lee, Woo-Kyoung;Park, Jong-Sang
    • BMB Reports
    • /
    • v.30 no.3
    • /
    • pp.211-215
    • /
    • 1997
  • A photoactive analog of ATP, 3'(2')-O-(p-azidobenzoyl)-adenosine 5-triphosphate (AB-ATP) was synthesized by chemically coupling N-hydroxysuccinimidyl-4-azidobenzoate (NHS-AB) and ATP. The utility of AB-ATP as an effective active-site-directed photoprobe was demonstrated using catalytic subunit of protein kinase A as a model enzyme. Photoincorporation of AB-ATP was saturated with apparent dissociation constant of $30{\mu}m$ and protected completely by $100{\mu}m$ of ATP. When the enzyme was covalently modified by photolysis in the presence of saturating amounts of photoprobe, about 60% inhibition of enzyme activity was observed. These results demonstrate that AB-ATP has potential application as a probe to characterize ATP-binding proteins including protein kinases.

  • PDF

Effect of ATP on Calcium Channel Modulation in Rat Adrenal Chromaffin Cells (흰쥐 부신 크로마핀 세포 칼슘통로 조절에 미치는 ATP의 효과)

  • Kim, Kyung Ah;Goo, Yong Sook
    • Progress in Medical Physics
    • /
    • v.25 no.3
    • /
    • pp.157-166
    • /
    • 2014
  • ATP in quantity co-stored with neurotransmitters in the secretory vesicles of neurons, by being co-released with the neurotransmitters, takes an important role to modulate the stimulus-secretion response of neurotransmitters. Here, in this study, the modulatory effect of ATP was studied in $Ca^{2+}$ channels of cultured rat adrenal chromaffin cells to investigate the physiological role of ATP in neurons. The $Ca^{2+}$ channel current was recorded in a whole-cell patch clamp configuration, which was modulated by ATP. In 10 mM $Ba^{2+}$ bath solution, ATP treatment (0.1 mM) decreased the $Ba^{2+}$ current by an average of $36{\pm}6%$ (n=8), showing a dose-dependency within the range of $10^{-4}{\sim}10^{-1}mM$. The current was recovered by ATP washout, demonstrating its reversible pattern. This current blockade effect of ATP was disinhibited by a large prepulse up to +80 mV, since the $Ba^{2+}$ current increment was larger when treated with ATP ($37{\pm}5%$, n=11) compared to the control ($25{\pm}3%$, n=12, without ATP). The $Ba^{2+}$ current was recorded with $GTP{\gamma}S$, the non-hydrolyzable GTP analogue, to determine if the blocking effect of ATP was mediated by G-protein. The $Ba^{2+}$ current decreased down to 45% of control with $GTP{\gamma}S$. With a large prepulse (+80 mV), the current increment was $34{\pm}4%$ (n=19), which $25{\pm}3%$ (n=12) under control condition (without $GTP{\gamma}S$). The $Ba^{2+}$ current waveform was well fitted to a single-exponential curve for the control, while a double-exponential curve best fitted the current signal with ATP or $GTP{\gamma}S$. In other words, a slow activation component appeared with ATP or $GTP{\gamma}S$, which suggested that both ATP and $GTP{\gamma}S$ caused slower activation of $Ca^{2+}$ channels via the same mechanism. The results suggest that ATP may block the $Ca^{2+}$ channels by G-protein and this $Ca^{2+}$ channel blocking effect of ATP is important in autocrine (or paracrine) inhibition of adrenaline secretion in chromaffin cell.