• 제목/요약/키워드: ATP hydrolysis

검색결과 60건 처리시간 0.028초

Deinococcus radiodurans RecA 단백질의 외가닥 DNA-의존성 ATPase 활성 분석 (Characterization of Single Stranded DNA-Dependent ATPase Activities of Deinococcus radiodurans RecA Protein)

  • 김종일
    • 미생물학회지
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    • 제43권4호
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    • pp.250-255
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    • 2007
  • Deinococcus radiodurans recA는 이 미생물의 방사선 저항성을 나타내는 표현형에 필수적이며 재조합성 DNA 수선 과정에 관여한다. 이 과정에서 RecA 단백질은DNA와 결합하여 반응의 활성 종인 RecA nucleoprotein 필라멘트를 형성한다. DNA-의존성 ATPase 활성과 함께, RecA 단배질의 외가닥 DNA 혹은 이중가닥 DNA와의 상호작용은 RecA 단백질이 관여하는 반응의 중심과정으로 이에 관한 분석을 시도하였다. D. radiodurans RecA 단배질은 DNA에 결합한 DNA-단백질 복합체만이 ATPase 활성을 나타내므로, ATP (혹은 dATP) 가수분해를 측정함으로써 RecA와 외가닥 DNA와의 상호작용 정도를 분석하였다. D. radiodurans RecA 단백질은 외가닥 DNA의 염기 구성의 이질성에 영향을 받았으며, homopolymer인 poly(dT)와의 상호작용에서 가장 높은 가수분해 활성을 보였다. Homopolymer인 합성 DNA-의존성 ATP 및 dATP의 가수분해는 pH 6.0과 9.0의 범위에서 다소 일정한속도로 일어났으며 최적 pH는 7.0과 7.5 사이였다. 외가닥 DNA-의존성 ATPase 활성은 염의 존재에 영향을 받아 KCl이 존재하면 다소 억제되나, K-glutamate가 존재하면 오히려 촉진되었다. RecA 단백질과 외가닥 DNA의 상호작용을 ATP 가수분해로 분석하였을 때 2 mM 이상의 magnesium 이온이 DNA 결합반응에 필요하였으며, 비교적 넓은 범위의 pH에서 외가닥 DNA와의 결합반응이 일어나며, 이러한 결합반응은 당량적인 비(1:3, RecA protein: DNA nucleotide)로 일어났다.

Dependence of RIG-I Nucleic Acid-Binding and ATP Hydrolysis on Activation of Type I Interferon Response

  • Yu Mi Baek;Soojin Yoon;Yeo Eun Hwang;Dong-Eun Kim
    • IMMUNE NETWORK
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    • 제16권4호
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    • pp.249-255
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    • 2016
  • Exogenous nucleic acids induce an innate immune response in mammalian host cells through activation of the retinoic acid-inducible gene I (RIG-I). We evaluated RIG-I protein for RNA binding and ATPase stimulation with RNA ligands to investigate the correlation with the extent of immune response through RIG-I activation in cells. RIG-I protein favored blunt-ended, double-stranded RNA (dsRNA) ligands over sticky-ended dsRNA. Moreover, the presence of the 5'-triphosphate (5'-ppp) moiety in dsRNA further enhanced binding affinity to RIG-I. Two structural motifs in RNA, blunt ends in dsRNA and 5'-ppp, stimulated the ATP hydrolysis activity of RIG-I. These structural motifs also strongly induced IFN expression as an innate immune response in cells. Therefore, we suggest that IFN induction through RIG-I activation is mainly determined by structural motifs in dsRNA that increase its affinity for RIG-I protein and stimulate ATPase activity in RIG-I.

식물엽의 ATP함량에 미치는 중금속(Al, Cd, Hg 및 Pb)의 영향 (Effects of Some Heavy Metals(Al, Cd, Hg, and Pb) on ATP Content in Plant Leaves)

  • 성민웅
    • Journal of Plant Biology
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    • 제22권4호
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    • pp.107-113
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    • 1979
  • The present study was carried out to estimate the comparative effects of hydroponic heavy metals (Al, Cd, Hg, and Pb) on ATP content in plant leaves grown with Hoagland solution under green house condition. The two plants, kidneybean(Phaseolus vulgaris L.) and buckwheat(Fagopyrum esculentum M nch), showed similar inhibitory effect of heavy metals on ATP content in order of Hg, Cd, Pb, and Al. But the overall inhibitory effect was greater in kidneybean than in buckwheat. The affinity of heavy metals, in vitro, toward the enzyme (luciferin-luciferase) is in order of Hg, Al, Cd, and Pb, similar to that toward ATP. The results showed that the inhibitory effect of heavy metals on ATP hydrolysis is mainly due to the coordination of heavy metals with enzyme than ATP.

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Real-Time Monitoring of Mitochondrial ATP Synthesis and Hydrolysis by Surface Infrared Spectroscopy

  • Yamaguchi, Ryo-Taro;Hirano-Iwata, Ayumi;Aonuma, Yuki;Yoshimura, Yuya;Shinohara, Yasuo;Kimura, Yasuo;Niwano, Michio
    • 한국진공학회:학술대회논문집
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    • 한국진공학회 2013년도 제44회 동계 정기학술대회 초록집
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    • pp.108-109
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    • 2013
  • Mitochondria play key roles in the production of cell's energy. Their dominant function is the synthesis of adenosine 5'-triphosphate (ATP) from adenosine diphosphate (ADP) and phosphate (Pi) through the oxidative phosphorylation. Evaluation of drug-induced mitochondrial toxicity has become increasingly important since mitochondrial dysfunction has recently been implicated in numerous diseases including cancer and diabetes mellitus. Mitochondrial functions have been monitored via oxygen consumption, mitochondrial membrane potential, and more importantly via ATP synthesis since ATP synthesis is the most essential function of mitochondria. Various analytical methods have been employed to investigate ATP synthesis in mitochondria, including high performance liquid chromatography (HPLC), bioluminescence technique, and pH measurement. However, most of these methods are based on destructive analysis or indirect monitoring through the enzymatic reaction. Infrared absorption spectroscopy (IRAS) is one of the useful techniques for real-time, label-free, and direct monitoring of biological reactions [1,2]. However, the strong water absorption requires very short path length in the order of several micrometers. Transmission measurements with thin path length are not suitable for mitochondrial assays because solution handlings necessary for evaluating mitochondrial toxicity, such as rapid mixing of drugs and oxygen supply, are difficult in such a narrow space. On the other hand, IRAS in the multiple internal reflection (MIR) geometry provides an ideal optical configuration to combine solution handling and aqueous-phase measurement. We have recently reportedon a real-time monitoring of drug-induced necrotic and apoptotic cell death using MIR-IRAS [3,4]. Clear discrimination between viable and damaged cells has been demonstrated, showing a promise as a label-free and real-time detection for cell-based assays. In the present study, we have applied our MIR-IRAS system to mitochondria-based assays by monitoring ATP synthesis in isolated mitochondria from rat livers. Mitochondrial ATP synthesis and hydrolysis were in situ monitored with MIR-IRAS, while dissolved oxygen level and solution pH were simultaneously monitored with O2 and pH electrodes, respectively. It is demonstrated that ATP synthesis and hydrolysis can be monitored by the IR spectral changes in phosphate groups in adenine nucleotides and MIR-IRAS is useful for evaluating time-dependent drug effects of mitochondrial toxicants.

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Nucleotide and Manganese Ion is Required for Chaperonin Function of the Hyperthermostable Group II Chaperonin α from Aeropyrum pernix K1

  • Jang, Kyoung-Jin;Bae, Yu-Jin;Jeon, Sung-Jong;Kim, Kyung-Hwa;Lee, Jung-Hee;Yea, Sung-Su;Oh, Sang-Taek;Jeong, Yong-Joo;Kim, Dong-Eun
    • Bulletin of the Korean Chemical Society
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    • 제28권12호
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    • pp.2261-2265
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    • 2007
  • Prevention of thermal aggregation of the denatured protein by the group II chaperonin from the aerobic hyperthermophilic crenarchaeon Aeropyrum pernix K1 (ApcpnA) has been investigated. ApcpnA exists as a homo-oligomer in a ring structure, which protects thermal aggregation of the chemically denatured bovine rhodanese at 50 oC. ApcpnA alone is not sufficient for chaperonin activity, but the chaperonin activity is greatly enhanced in the presence of manganese ion and ATP. Compared to the mesophilic chaperonin GroEL/GroES, ApcpnA is more activated at a higher temperature and protects the aggregation-prone unfolded state of the denatured rhodanese from thermal aggregation. Binding of ATP is sufficient for ApcpnA to perform the chaperonin function in vitro, but hydrolysis of ATP is not necessarily required. We propose that utilization of Mn2+ and adenosine nucleotide regardless of ATP hydrolysis may be one of peculiar properties of archaeal chaperonins.

Escherichia coli내의 ATP-dependent Clp효소의 ATPase 활성 연구 (Properties of ATPase Activity of ATP-dependent Clp Protease in Escherichia coli)

  • 김승호
    • 한국미생물·생명공학회지
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    • 제21권1호
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    • pp.30-35
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    • 1993
  • E.coli에서 발견된 ATP-dependent 효소인 Clp효소 중에서 Clp A의 ATPase 활성에 대한 영향을 검토하였다. Clp효소의 limiting amount으로 나타난 specific 활성은 일정하게 증가하는 효소의존성을 보였다. ATPase 활성을 나타내고 있는 ClP A는 casein에 의하여 활성화되어지며 2분자의 ATP가 결합하고 ATPase 활성을 나타내기 위한 ATP의 분해는 Clp효소의 단백질 분해 활성에 필요하다.

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ATP and GTP Hydrolytic Function of N-terminally Deleted Annexin I

  • Hyun, Young-Lan;Park, Young-Min;Na, Doe-Sun
    • BMB Reports
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    • 제33권4호
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    • pp.289-293
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    • 2000
  • Annexin I is a 37 kDa member of the annexin family of calcium-dependent phospholipid binding proteins. Annexin I plays regulatory roles in various cellular processes including cell proliferation and differentiation. Recently we found that annexin I is a heat shock protein (HSP) and displays a chaperone-like function. In this paper we investigated the function of annexin I as an ATPase using 1 to 32 amino acids deleted annexin I (${\Delta}-annexin$ I). ${\Delta}-Annexin$ I hydrolyzed ATP as determined by thin layer chromatography. The ability of ATP hydrolysis was inhibited by ADP, GTP and GDP, but not by the AMP, GMP and cAMP. In view of the ATP hydrolyzing function of HSP, the results support the function of annexin I as a HSP.

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수축중인 근육막에서 actin과 myosin 결합의 구조변화에 관한 연구 (A Study On Structure Change of Binding Actin and Moysin On The Contracting Muscle Membrane)

  • 김덕술;박근호
    • 한국응용과학기술학회지
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    • 제25권3호
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    • pp.380-387
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    • 2008
  • The effects of the applied stretch and MgADP binding on the structure of the actin and myosin cross-bridges in rabbit fibers in the rigor state have been investigatedwith improved resolution by x-ray diffraction using synchrotron radiation. To clarify the structure of the ATP hydrolysis intermediates formed by actin and myosin cross-bridges,the effects of various phosphate analogs in the of MgADP on the structure of the thin and thick filaments in glycerinated rabbit muscle fibers in the rigor state investigated by x-ray diffraction with a short exposure time using synchrotron radiation. These results strongly suggest that when MgADP and phosphate analogs such as metallofluorides(BeF3 and AlF4)and vanadate(VO4(Vi)) were added the rigor fibers in the presence of the ATP-depletion backup system, the intensities of the actin-based layer lines were markedly weakened. We found that the intensity of the 14.5 nm-based meridional reflections increase by 20-50% when phosphate analogs such as metallofluorides(BeF3 and AlF4) and vanadate(VO4(Vi)) was added to the rigor muscle.

정어리액젓의 숙성 중 성분변화 (Changes of Components in Salt-Fermented Sardine, Sardinops melanostictus Sauce during Fermentation)

  • 임영선;유병진;조영제
    • 한국수산과학회지
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    • 제35권6호
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    • pp.666-670
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    • 2002
  • 정어리 액젓을 재래식 방법으로 18개월 동안 숙성시키면서 2$\~$3개월 간격으로 성분변화에 대하여 조사하였다. 정어리 육의 가수분해도는 숙성 5개월까지는 $44\%$로 큰 폭으로 증가하였으나, 그 이후에는 분해속도가 둔화되어 숙성 18개월 후에는 약 $64\%$의 질소화합물이 육으로부터 액으로 이행되었다. 액젓 중의 총 질소 및 아미노태 질소함량, 그리고 ATP 관련물질 총량은 숙성기간에 비례하여 증가하였으며, 숙성 6개월 이후 ATP 관련물질은 거의 대부분 ($75\~87\%$)이 Hx와 요산이었다. HXR+HX 함량과 요산량이 교차하는 숙성 13.9개월 부근은 $61\%$의 높은 분해율을 보여 경제적인 출하시점으로 판단된다. 18개월간 숙성시킨 정어리 액젓의 유리아미노산 총량은 6,870.7mg/100mL로 원료 육 총 아미노산 함량 (23,249mg1100g)의 약 $30\%$ 정도였으며, 주요 아미노산은 glutamic acid ($18.2\%$), aspartic acid ($10.4\%$), lysine ($8.7\%$), alanine ($8.5\%$), threonine ($6.1\%$) 등의 순이었다.

중급 지방산 항진균 활성과 진균의 Plasma membrane H+-ATPase에 대한 저해작용 (Antifungal Activity of Medium-chain Saturated Fatty Acids and Their Inhibitory Activity to the Plasma Membrane H+-ATPase of Fungi)

  • 이상화;김창진
    • 한국미생물·생명공학회지
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    • 제27권5호
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    • pp.354-358
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    • 1999
  • In order to know the antifungal characteristics of saturated fatty acids having 6 to 12 carbons, their minimum inhibitory concentrations (MICs) and minimum fungicidal concentrations (MFCs) were estimated against Saccharomyces cerevisiae. Fatty acids from C6 to C11 exhibited increasing activity with chain length, but C12 fatty acid did not show activity at all. In relation to antifungal modes of actions, fatty acids investigated showed on inhibitory activity toward the plasma membrane H+-ATPase of Saccharomyces cerevisiae. Their inhibitions to the glucose-induced acidification and ATP hydrolysis caused by the proton pump were found to be in common wiht antifungal activities. At the test concentration of 1mM, hexanoic acid (C6) showed the lowest inhibition of about 30%, while undecanoic acid(C11) showed the strongest inhibition of over 90%. In addition, as seen with antifungal activity, the inhibitory activity of dodecanoic acid (C12) was suddenly reduced to less than 50%.

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