• 제목/요약/키워드: K-ATPase protein

검색결과 229건 처리시간 0.034초

Adipic acid 저항성 변이주 Leuconostoc paramesenteroides의 내산성 특성 (Properties of Acid Tolerance of the Adipic acid-resistant Mutant of Leuconostoc paramesenteroides)

  • 이중근;이홍석;김영찬;주현규;이시경;강상모
    • 한국식품과학회지
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    • 제32권2호
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    • pp.431-438
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    • 2000
  • Adipic acid에 대해 저항성을 갖도록 변이된 Leuconostoc paramesenteroides (ANaP100) 균주의 내산성 특성을 조사하기 위하여 수소이온 투과도, $H^+-ATPase$ 활성, $Mg^{++}$ 해리도, 원형질막의 지방산 조성을 지표로 삼아 야생균주(LPw)와 비교하였다. 수소이온 투과도의 경우 pH 5에서 $t_{1/2}$ 값이 LPw는 4.3분, ANaP100은 4.8분으로서 변이균주가 다소 내산성이 높았으며 $H^+-ATPase$ 활성은 maximal activity가 Leu. paramesenteroides는 모두 pH 6.0에서 가장 높았고 LPw는 0.59 unit/mg protein, 변이균주는 0.63 unit/mg protein으로서 ANaP100이 LPw보다 활성이 높았다. 세포막의 산 손상(acid damage)에 의한 $Mg^{++}$ 해리도에서도 pH 4.0에서 2시간 경과 후 LPw는 52.2%, ANaP100은 27.3%로서 LPw에 비해 약 1/2가량 $Mg^{++}$이 적게 유출되어 산에 의한 세포막의 손상이 적었다. 원형질막의 지방산 조성은 ANaP100가 $C_{18:1}$은 감소하고, $C_{19:0,\;cyclo}$는 증가하여 내산성이 증대되었으며 또한 adipic acid 첨가시에도 LPw에 비해 우수한 증식을 보였다. 따라서 ANaP100이 LPw에 비해 내산성이 증가되었으며 adipic acid 저항성도 함유하는 것이 확인되었다.

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Suppression of the ER-Localized AAA ATPase NgCDC48 Inhibits Tobacco Growth and Development

  • Bae, Hansol;Choi, Soo Min;Yang, Seong Wook;Pai, Hyun-Sook;Kim, Woo Taek
    • Molecules and Cells
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    • 제28권1호
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    • pp.57-65
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    • 2009
  • CDC48 is a member of the AAA ATPase superfamily. Yeast CDC48 and its mammalian homolog p97 are implicated in diverse cellular processes, including mitosis, membrane fusion, and ubiquitin-dependent protein degradation. However, the cellular functions of plant CDC48 proteins are largely unknown. In the present study, we performed virus-induced gene silencing (VIGS) screening and found that silencing of a gene encoding a tobacco CDC48 homolog, NgCDC48, resulted in severe abnormalities in leaf and shoot development in tobacco. Furthermore, transgenic tobacco plants (35S:anti-NgCDC48), in which the NgCDC48 gene was suppressed using the antisense RNA method, exhibited severely aberrant development of both vegetative and reproductive organs, resulting in arrested shoot and leaf growth and sterile flowers. Approximately 57-83% of 35S:anti-NgCDC48 plants failed to develop mature organs and died at early stage of development. Scanning electron microscopy showed that both adaxial and abaxial epidermal pavement cells in antisense transgenic leaves were significantly smaller and more numerous than those in wild type leaves. These results indicate that NgCDC48 is critically involved in cell growth and development of tobacco plants. An in vivo targeting experiment revealed that NgCDC48 resides in the endoplasmic reticulum (ER) in tobacco protoplasts. We consider the tantalizing possibility that CDC48-mediated degradation of an as-yet unidentified protein(s) in the ER might be a critical step for cell growth and expansion in tobacco leaves.

Effects of High Glucose Levels on the Protein Kinase C Signal Transduction Pathway in Primary Cultured Renal Proximal Tubule Cells

  • Han, Ho-Jae;Kang, Ju-Won;Park, Kwon-Moo
    • The Korean Journal of Physiology
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    • 제30권2호
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    • pp.257-267
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    • 1996
  • Diabetes mellitus is associated with a wide range of pathophysiologic changes in the kidney. This study was designed to examine the mechanisms by which glucose modulates the expression of polarized membrane transport functions in primary cultured rabbit renal proximal tubule cells. Results are as follows: The rate of 30 minute $Rb^{+}$ uptake was significantly higher($137.76{\pm}5.40%$) in primary renal tubular cell cultures treated with 20 mM glucose than that of 5 mM glucose. Not the level of mRNA for the ${\alpha}$ subunit of Na, K-ATPase but that of ${\beta}$ subunit was elevated in primary cultures treated with high glucose. The initial rate of methyl-${\alpha}$-D-glucopyranoside(${\alpha}$-MG) uptake was significantly lower($71.91{\pm}3.02%$) in monolayers treated with 20 mM glucose than that of 5 mM glucose. There was a tendency of an increase in phlorizin binding site in cells treated with 5 mM glucose. However, 3-O-methyl-D-glucose(3-O-MG) uptake was not affected by glucose concentration in culture media. TPA inhibited $Rb^{+}$ uptake by $63.61{\pm}1.94\;and\;45.80{\pm}1.36%$ and ${\alpha}$-MG uptake by $48.54{\pm}3.69\;and\;41.87{\pm}6.70%$ in the cells treated with 5 and 20 mM glucose, respectively. Also TPA inhibited mRNA expression of Na/glucose cotransporter in cells grown in 5mM glucose medium. cAMP significantly stimulated ${\alpha}$-MG uptake by $114.65{\pm}5.70%$ in cells treated with 5mM glucose, while it did not affect ${\alpha}$-MG uptake in cell treated with 20 mM glucose. However, cAMP inhibited $Rb^{+}$ uptake by $76.69{\pm}4.16\;and\;66.87{\pm}2.41%$ in cells treated with 5 and 20 mM glucose, respectively. In conclusion, the activity of the renal proximal tubular Na,K-ATPase is elevated in high glucose concentration. In contrast, the activity of the Na/glucose cotransport system is inhibited. High glucose may in part affect the activity of the Na,K-ATPase and the Na/glucose cotransport system by controlling the protein kinase C and/or A signal transduction pathway in primary cultured renal proximal tubule cells.

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건조 및 저장조건에 따른 쌀 저장단백질의 변화양상 (Changes of Rice Storage Proteins Affected by Dry and Storage Temperature)

  • 신평균;장안철;홍성창;이기상;이금희;이용복
    • 한국환경농학회지
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    • 제27권4호
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    • pp.456-459
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    • 2008
  • Quality of rice grain changes during dry storage with internal physiological changes and external injury by organism. Storage rice changes by condition with respiration via variable temperature, hydrolysis enzyme reaction, lipid peroxidation occurs with change of palatability. During dry storage, physiological change with protein variation pattern was examined by image analysis on proteomic technology. Analysis revealed that protein activity had no change store at room temperature and store at $40^{\circ}C$, but decreased store at $60^{\circ}C$. Analysis of variable hydrophobic protein pattern revealed that protein activity of beta-tubulin, protein disulfide isomerase, vacuolar ATPase b subunit, globulin was not significantly decreased all dry and store condition. However, heat shock protein 70, and glutathione transferase was significantly decreased when rice dried at $60^{\circ}C$ compared with room temperature and $40^{\circ}C$ dry condition.

Identification of antigenic proteins of lymphocystis disease virus (LCDV) by MALDI-TOF mass spectrometry

  • Chung, Chang-Kyun;Kim, Byung-Gwan;Jung, Myung-Hwa;Jung, Sung-Ju
    • 한국어병학회지
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    • 제28권3호
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    • pp.133-143
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    • 2015
  • The antigenic proteins of Lymphocystis disease virus (LCDV) from tumors of olive flounder, Paralichthys olivaceus, are described following characterization by mass spectrometry. In SDS-PAGE, predominant protein bands were observed at 114, 88, 70, 54, 52, 47, 42 and 24 kDa. Western blot analysis showed that antisera reacted strongly at molecular weights of 114, 67 and 54 kDa, and reacted weakly at molecular weights of 74, 70, 36, 24 and 22 kDa. In the identification of LCDV antigenic proteins by matrix-assisted laser desorption ionization (MALDI) TOF mass spectrometry, 10 of 14 excised bands consisted mostly of proteins with amino acid sequences that matched LCDV-C (lymphocystis disease virus isolate China) ORFs. Strong antigens with molecular weights of 114, 67 and 54 kDa were identified as LDVICp236 (chromosome segregation ATPase), LDVICp033 (membrane bound metallopeptidase) and LDVICp157 (hypothetical protein), respectively. Minor antigens with molecular weights of 70, 36, 24 and 22 kDa proteins were identified as LDVICp160 (acetyl-coA hydrolase), LDVICp213 (hypothetical protein), LDVICp039 (hypothetical protein) and LDVICp213 (hypothetical protein). However, the major capsid protein (LDVICp043) did not react with the polyclonal antibody.

Physiological Relevance of Salt Environment for in vitro recA System

  • Kim, Jong-Il
    • Journal of Microbiology
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    • 제37권2호
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    • pp.59-65
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    • 1999
  • RecA protein can promote strand assimilation, homologous pairing, and strand exchange. All these reactions require DNA-dependent ATP hydrolysis by recA protein, and the activities of recA protein are affected by the ionic environment. In this experiment, DNA-dependent ATPase activity showed different sensitivity to anionic species. ATP hydrolysis and strand exchange were relatively sensitive to salt in the reactions with NaCl, strongly inhibited at 100 mM NaCl. However, the inhibition by sodium acetate or sodium glutamate was not observed at 50∼100 mM concentration. Addition of sodium glutamate to the standard reaction condition increased the apparent efficiency of ATP hydrolysis during strand exchange. The condition including 50∼100 mM sodium-glutamate might be similar to the physiological condition.

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해산복족류(海産腹足類)의 Paramyosin의 분리(分離) 및 그 특성(特性)에 관(關)한 연구(硏究) (Isolation and Characterization of Paramyosins of Marine Gastropods)

  • 변재형
    • 한국식품영양과학회지
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    • 제2권1호
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    • pp.1-21
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    • 1973
  • The muscle of abalone, Notohaliotis discus (REEVE), and top-shell, Turbo cornutus Solander, were examined for protein composition. Then paramyosins which are known as one of the important structural protein of the muscle fibrils were isolated from the both muscle and their physico-chemical properties such as solubility, salting-out behaviour, intrinsic viscosity, ATPase activity, etc. involving amino acid composition and N-terminal amino acid residues were investigated to elucidate phylogenie characteristics more intensively from the viewpoint of comparative biochemistry. The analysis of protein composition resulted in the following estimations: abalone muscle; water-soluble protein of 22 %, salt-soluble protein, 34%, alkali-soluble protein, 20%, and stroma protein, 24%, and top-shell muscle; water-soluble protein of 16%, salt-soluble protein, 30%, alkali-soluble protein, 29%, and stroma protein, 25%, respectively. It is demonstrated in sedimentation analysis that paramyosin and myosin-actomyosin account for approximately 65% and 35% of the salt-soluble protein of abalone, and that the composition of both sediments in top-shell was approximately 70% and 30%, respectively. The ultracentrifugally homogenous paramyosins isolated essentially according to Bailey's ethanol-dried method from both of the muscle showed a $S^{\circ}_{20,w}$ of 3. 14s for abalone and a $S^{\circ}_{20,w}$ of 3.50s for top-shell. The both paramyosins were commonly rich in arginine, aspartic acid, and glutamic acid, while scarcely contained proline and tryptophan, in rough accord with the other paramyosins thus far reported. It is clear that these gastropod paramyosins showed of having the characteristic N-terminal amino acid residues such as N-aspartic acid, N-valine, N-serine, and N-threonine in common. The abalone paramyosin completely salted in with KCl beyond $0.35{\mu}$ and the top-shell paramyosin beyond $0.30{\mu}$. The abalone paramyosin was salted-out between 18% and 30% saturation of ammonium sulphate and the top-shell paramyosin between 22% and 29% saturation. The intrinsic viscosities at abalone and top-shell paramyosins at $25^{\circ}C$ were estimated respectively to be 3.1 dl/g and 2.6 dl/g showing somewhat higher than the values for some other paramyosins from lamellibranchs. In regard with the ATPase activity, the para myosin specimens did not exhibit any significant activity over through the pH conditions of 5 to 9.5. irrespective of the presence of $Ca^{++}$ or $Mg^{++}$. So was the case with the abalone paramyosin prepared by a slightly modified Bailey's wet-extraction method.

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한우 난포란의 체외성숙 배지와 세포질 내의 단백질 변화 (The Changes of Quantity and Quality of Proteins in Medium and Cytoplasm during In Vitro Maturation of Bovine Oocytes)

  • 박용수;박현정
    • Reproductive and Developmental Biology
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    • 제29권3호
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    • pp.187-191
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    • 2005
  • 본 연구는 한우 난포란이 체외성숙된 환경의 변화를 단백질 측면으로부터 검토하기 위하여 체외성숙 배지와 세포질내 단백질 변화와 종류를 검토하였다. 그 결과 배지 내의 단백질 발현량은 배양 4.5시간째까지 감소하였고, 배양 13.5시간째까지는 변화가 없었다. 그러나 배양 $13.5\~18$시간 사이에 증가한 후 배양 18시간 이후에 다시 감소하는 경향이었다. 세포질내의 단백질 발현량은 배양 4.5시간째까지 증가하였고 배양 9시간째까지 급격히 감소하였다. 배양 9시간째부터 18시간째 까지는 단백질 발현량이 유사한 경향이었으나 배양 18시간째부터 24시간째까지 다시 증가하였다. 한편 체외성숙한 배지와 세포질을 2차원 전기영동하여 각각 298개 및 35개의 단백질 spot을 확인하였고, 그 중 배지에서는 28개, 세포질에서는 5개의 spot이 유의적인 변화를 확인하였다. 이들 spot 대한MALDI-TOP분석으로 배지와 세포질에서 각각 8개 및 1개의 단백질을 동정하였다. 그 종류는 aldose reductase, alpha enolase, apolipoprotein A-1 precursor, 43M1a collectin precursor, heat shock 27kDa protein, plasminogen activator inhibitor-1 precursor, thrombospondin 1 transitional endoplasmic reticulum ATPase 및 $\beta$-tubulin이었다.

Proteome Analysis of the Young Spikelets of Photoperiod-Sensitive Rice Mutant Treated in Different Photoperiods

  • Pandeya, Devendra;Song, You-Chun;Kim, Sung-Su;Suh, Hak-Soo;Kang, Sang-Gu
    • 한국작물학회지
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    • 제52권3호
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    • pp.281-288
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    • 2007
  • Photoperiod sensitive genetic male sterile (PGMS) rice is sterile mutant controlled by photoperiod. A PGMS mutant 920S was sterile grown under long-day (LD) photoperiod (14 h light/10 h dark) but fertile grown under short-day (SD) photoperiod (10 h light/14 h dark). Proteome analysis revealed that 12 protein spots were differentially expressed in the spikelets of 920S plants either treated with LD or SD photoperiod. Among these proteins, three proteins including chlorophyll a/b binding protein, vacuolar ATPase ${\beta}-subunit,\;{\alpha}-tubulin$ and an unknown protein were more than three-fold abundant in the spikelet of the SD-treated plants than those of the LD-treated plants. On the other hand, eight proteins including acetyl transferase, 2, 3- biphosphoglycerate, aminopeptidase N, pyruvate decarboxylase, 60S acidic ribosomal protein and three unknown protein spots were more abundant in the spikelets of the LD-treated plants than those of the SD-treated plants. The results suggest that the observed proteins may be involved in sterile or fertile pollen development under LD or SD photoperiod respectively in the PGMS mutant rice.

전분가수분해물의 어육단백질 동결변성 방지효과 및 작용기구 1. 옥수수전분가수분해물의 어육단백질에 대한 동결변성 방지효과 (Cryoprotective Effect and Mechanism of Corn Starch Enzyme Hydrolysates on Fish Protein 1. Cryoprotective Effect of Corn Starch Enzyme Hydrolysates on Fish Protein)

  • 이강호;정병천;홍병일
    • 한국수산과학회지
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    • 제31권6호
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    • pp.823-828
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    • 1998
  • 명태 actomyosin 용액에 대한 옥수수전분 가수분해물의 동결 보호 효과는 전기영동상 패턴, 용해도 및 $Ca^{2+}$-ATPase 활성 변화로 알아보았다. 먼저 전기영동상 패턴에서 0일차와 저장 30일차를 비교하였을 때, control의 경우 저장온도에 관계없이 각 band들의 변화가 수반되었고, 특히 MHC의 변화가 두드러졌다. Sucrose 첨가구의 경우, 저장온도에 상관없이 band상의 변화가 거의 없는 것으로 나타났다. 전분가수분해물이 첨가된 시료들에서는 온도 의존성이 현저하여 $-5^{\circ}C$에서 저장한 시료들은 MHC band가 나타나지 않았고 반면에 $-20^{\circ}C$에서는 저장 30일 째에도 0일차와 거의 같은 양상을 보였다. 시료들을 $-5^{\circ}C$에서 저장할 때 용해도 변화는 sucrose의 경우 저장 30일 후에도 $90\%$ 이상의 높은 값을 유지한 반면에 나머지 시료들은 저장과 함께 급속히 감소하였다 $-20^{\circ}C$에서는 sucrose, D.E. 10, 15 및 20을 함유한 시료들은 약 $90\%$ 정도를 나타내었다. 잔존 활성 변화에서도 용해도 변화와 거의 유사한 경향을 나타내었다. 이상에서 살펴보았듯이 명태 actomyosin 용액의 동결변성에 대한 옥수수전분가수분해물들의 보호효과는 저장온도에 따라 많은 차이를 나타내었고, 특히 D.E. 10의 경우는 기존의 동결변성 방지제인 sucrose와 비교하여 $-12^{\circ}C$ 이하에서는 거의 같은 효과를 나타내는 것을 알았고, 저감미, 저열량의 동결변성 방지제로서 가능한 것으로 사료된다.

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