• Title, Summary, Keyword: AB-ATP

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

  • Shin, Seung-Jin;Lee, Woo-Kyoung;Park, Jong-Sang
    • BMB Reports
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    • v.30 no.3
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    • pp.211-215
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    • 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.

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Protectors of Oxidative Stress Inhibit AB(1-42) Aggregation in vitro

  • Kong, Byung-Mun;Ueom, Jeong-Hoon;Kim, In-Kyung;Lim, Dong-Yeol;Kang, Jong-Min;Lee, Kyung-Hee
    • Bulletin of the Korean Chemical Society
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    • v.23 no.12
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    • pp.1773-1777
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    • 2002
  • Reactive oxygen species(ROS) have been investigated to have pivotal roles on amyloidogenecity of $\beta-amyloidpeptide(A\beta)$, the major component of senile plaques in Alzheimer's disease(AD) brain. Addition of radical scavengers is one of the on-going strategies for therapeutic treatment for AD patients. Hsp104 protein including two ATP binding sites from Saccharomyces cerevisiae, as a molecular chaperone, was known to function as a protector of ROS generation when exposed to oxidative stress in our previous study. This observation has led us to investigate Hsp104 protein as a molecular mediator of $A{\beta}$ aggregation in this study. We have developed a new way of expression for Hsp104 protein using GST-fusion tag. As we expected, formation of $A{\beta}$ aggregate was protected by wild type Hsp104 protein, but not by the two ATP-binding site mutant, based on Thioflavin-T fluorescence. Interestingly, Hsp104 protein was observed to keep $A{\beta}$ from forming aggregates independent of ATP binding. On the other hand, disaggregation of $A{\beta}$ aggregates by wild type Hsp104 was totally dependent on the presence of ATP. On the other hand, mutant Hsp104 with two ATP binding sites altered exhibited no inhibition. Another effective antioxidant, hydrazine analogs of curcumin were also effective in $A{\beta}$ fibrilization as protectors against oxidative stress. Based on these observations we conclude that Hsp104 and curcumin derivatives, as protectors of oxidative stress, inhibit $A{\beta}$ aggregation in virto and can be candidates for therapeutic approaches in cure of some neurodegenerative disease.

Comparison Between ELISA and Gel-filtration Assay for the Guantitation of Airway Mucins

  • Shin, Chan-Young;Kang, Suk-Jo;Kim, Kwang-Chul;Ko, Kwang-Ho
    • Archives of Pharmacal Research
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    • v.21 no.3
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    • pp.253-259
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    • 1998
  • In this study, we developed immunoassay methods for the more convenient and effective detection of rat tracheal mucin and the results were compared with those of [$3^H$]glucosamine based gel-filtratioh method. A monoclonal anti-rat tracheal mucin antibody, mAbRT03, which specifically recognizes rat tracheal mucins, was used throughout in this study. To induce mucin secretion, varying concentrations of ATP (0-2 mM) were applied to the primary rat tracheal surface epithelial (RTSE) cell culture which had been metabolically radiolabeled with [$3^H$]glucosamine and the secretion of mucin was analyzed both by the immunoassay and the gel-filtration chromatography methods. For the immunoassay, the following two procedures were employed. 1) Simple ELISA; the culture spent media were directly coated onto the assay plate and the immunoreactivity with mAbRT03 was assessed from the standard curve generated with the purified rat mucin. 2) Inhibition ELISA; A known amount of the purified rat mucin was coated onto the assay plate and then ATP-stimulated culture spent media were added to inhibit the immunorelitivity with mAbRT03. The contents of mucin in the sample were calculated from the standard inhibition curve which was generated with the purified rat mucin. The assay results obtained from the immunoassays were identical with those from the gel-filtration methods. The present result indicates that ELISA can be substituted for the laborious, time-consuming gel-filtration assay in studying the regulation of airway mucin release from cultured airway epithelial cells.

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The Development of Integration Electronic Block System for Maintenance Efficiency on Railway Wayside Signalling System (철도 선로변 신호설비 유지보수 효율화를 위한 집중형 전자폐색제어장치 개발)

  • Baek, Jong-Hyen;Jo, Hyun-Jeong;Kim, Yong-Kyu
    • Journal of the Korea Academia-Industrial cooperation Society
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    • v.13 no.9
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    • pp.4171-4176
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    • 2012
  • The conventional block systems for railway signaling currently in operation in Korea have not been electrified or integrated and therefore there have been difficulties in terms of construction and maintenance. Two independent systems have been installed for ABS and LEU of ATP system (for speeded-up lines), although they deal with the same signaling information at the same location in order to control the trains. In these conventional ABS and LEU, a number of duplicate modules are installed in each device including lamp detection units and power supply units and it results increased manufacturing costs and maintenance efforts. This paper deals with the prototype development of integrated electronic block system with CPU-based digital control methods in order to overcome the limitations of the conventional ABS. The suggested system is the integration of the conventional ABS with LEU of ATP and it is also applicable for the non-ATP sections as well.

Pharmacological studies on aggressive behavior induced by lesions of the nucleus accumbens septi in rats

  • Lee, Soon-Chul;Ueki, Showa
    • Archives of Pharmacal Research
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    • v.9 no.3
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    • pp.169-174
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    • 1986
  • Bilateral lesion of nucleus accumbens septi (N, AB), one of the mesolimbic nuclei, resulted in hyperirritability and muricide including mouse eating behavior in rats. The effects of various drugs on hyperirritability and muricide induced by NAB lesion were investigated in rats. Hyperirritability in NAB rats were significantly reduced by L-DOPA L-5-HTP major and minor tranquilizers but not reduced by MA, ATP and imipriamine-like antidepressants. On the other hand, muricide in NAB rats was significantly suppressed by L-DOPA, L-5-HTP, major and minor tranquilizers, furthermore, selectively suppressed by MA, ATP and antidepressants. These results suggested that the neural mechanism for inducing muricide is distinct from for hyperirritability in NAB rats, and that muricide in NAB rats is resulted from the increasing of cholinergic activity and reduction of dopaminergic and serotonergic activity.

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Cyanobacterial bioreporters for detection of heavy metals, herbicide, and antibiotics (중금속, 제초제 및 항생제 검출용 남세균 유래 바이오 리포터)

  • Kim, Soo-Youn;Jeong, Won-Joong;Suh, Kye-Hong;Liu, Jang-Ryol;Park, Youn-Il
    • Journal of Plant Biotechnology
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    • v.35 no.2
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    • pp.141-145
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    • 2008
  • In this study, glucose-inducible intergenic sequences were used to generate bioreporters of the cyanobacterium Synechocystis sp. PCC 6803 that could monitor environmental pollutants. Luciferase genes LuxAB from the marine bacterium Vibrio fischeri under the control of glucose-inducible intergenic seqeucens of eight genes (atpI, ndbA, ctaD1, tkt, pgi, pdh, ppc, and cydA) were successfully expressed in the cyano-bacterial transformants, showing 5-25 fold increases in biolumeniscence upon exposure to glucose. In addition, glucose-inducible cyanobacterial bioreporters were very sensitive to various chemicals such as heavy metals ($Hg^{2+}$, $Cu^{2+}$, $Zn^{2+}$), electron transport inhibitors (DCMU, DBMIB, $CN^-$), and antibiotics (chloramphenicol and rifampicin). These glucose-inducible cyanobacterial bioreporters would be useful to develop biosensors for rapid screening of environmental samples.

The Study on Centralization & Electronic for Maintenance Efficiency of Ground Signaling System (지상신호설비의 유지보수 효율화를 위한 집중화 및 전자화 연구)

  • Baek, Jong-Hyen;Kim, Yong-Kyu;Lee, Kang-Mi
    • Journal of the Korea Academia-Industrial cooperation Society
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    • v.11 no.8
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    • pp.2983-2988
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    • 2010
  • The train control system used in Gyeongbu-line is classified in ATC, IXL and CTC. Domestic railway signaling systems are being developed by electrification. In these systems the electrification of interlocking reaches 57% and the safety equipments of railway crossings in trackside devices have completed their development into an integrated system. Block systems of all the existing sections have not yet electrified and integrated so that they need a number of complement in terms of construction and maintenance. For ABS currently used in existing domestic lines, and LEU being installed in Gyeongbu and Honam lines, although a train is controlled by the signaling information of the same train in the same location, the system is separately installed so that the same information is separately divided and transmitted at the each distinct system. Therefore, in the conventional ABS and LEU, there are a lot of duplicate installed compartments such as lamp detection and a power supply unit. Hence, we have a lot of problems: for maintenance, a lot of manpower and costs need to be invested and the overall manufacturing costs get higher, as well as the construction costs by duplicate. Therefore, this paper suggest design to develop an integrated electronic Block Control Unit by the integration of the currently used ABS, and communication and electronic technology. We are to monitor and manage the block systems in the corresponding station by integrating. And we are to transmit information together with LEU, which is an ATS wayside transmitter.

A NEWLY DEVELOPED CONTINUOUS TOXICITY TEST SYSTEM USING A LUMINOUSLY MODIFIED TERRESTRIAL BACTERIUM

  • Cho, Jang-Cheon;Lee, Kyu-Ho;Lee, Dong-Hun;Jahng, Deok-Jin;Park, Han-Oh;Kim, Sang-Jong
    • Proceedings of the Korean Society for Applied Microbiology Conference
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    • pp.108-113
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    • 2000
  • Freshwater borne bacteria transformed with luxAB-containing plasmid were optimized for the toxicity tests of various organic carbons and heavy metals. The EC$\sub$50/ values obtained from tests using the most sensitive bacterium to toxicants, YH9-RC, revealed to be much less than those from the Microtox$\^$/. In addition, some physiological characteristics of this bacterium under the toxic stress conditions such as potential bioluminescence, specific growth rate, and intracellular ATP contents, reproducibly and reliably correlated to the toxicity of the chemicals exposed. The higher concentrations of COD in wastewater samples, the lower EC$\sub$50/ values, therefore the developed toxicity test was found to be easily applicable to the toxicity test for wastewater samples and effluents. The conditions for constructing 384-multiwell plate containing freeze-dried bacterium were also optimized through the addition of 0.16 M trehalose before freeze-drying. Consequently, the advanced test system featuring a continuous measurement of the toxicity, an automated real-time monitoring of its results, and an alerting function was designed and constructed in combination with the microbiological, mechanical, and electronic compartment.

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Characterization of a PyrR-deficient Mutant of Bacillus subtilis by a Proteomic Approach (프로테옴 분석에 의한 Bacillus subtilis PyrR 돌연변이체의 특성)

  • Seul, Keyung-Jo;Cho, Hyun-Soo;Ghim, Sa-Youl
    • Microbiology and Biotechnology Letters
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    • v.39 no.1
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    • pp.9-19
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    • 2011
  • The Bacillus subtilis pyrimidine biosynthetic (pyr) operon encodes all of the enzymes for the de novo biosynthesis of Uridine monophosphate (UMP) and additional cistrones encoding a uracil permease and the regulatory protein PyrR. The PyrR is a bifunctional protein with pyr mRNA-binding regulatory funtion and uracil phosphoribosyltransferase activity. To study the global regulation by the pyrR deletion, the proteome comparison between Bacillus subtilis DB104 and Bacillus subtilis DB104 ${\Delta}$pyrR in the minimal medium without pyrimidines was employed. Proteome analysis of the cytosolic proteins from both strains by 2D-gel electrophoresis showed the variations in levels of protein expression. On the silver stained 2D-gel with an isoelectric point (pI) between 4 and 10, about 1,300 spots were detected and 172 spots showed quantitative variations in which 42 high quantitatively variant proteins were identified. The results showed that production of the pyrimidine biosynthetic enzymes (PyrAA, PyrAB, PyrB, PyrC, PyrD, and PyrF) were significantly increased in B. subtilis DB104 ${\Delta}$pyrR. Besides, proteins associated carbohydrate metabolism, elongation protein synthesis, metabolism of cofactors and vitamins, motility, tRNA synthetase, catalase, ATP-binding protein, and cell division protein FtsZ were overproduced in the PyrR-deficient mutant. Based on analytic results, the PyrR might be involved a number of other metabolisms or various phenomena in the bacterial cell besides the pyrimidine biosynthesis.