• Title/Summary/Keyword: ALPase

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생쥐 부정소 정자의 성숙과정에서 Phosphatase 활성도 변화 (Changes in Phosphatase Activities of Mouse Epididymal Spermatozoa during Maturation)

  • 김문규;윤현수;김종흡;김성례
    • 한국동물학회지
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    • 제33권1호
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    • pp.70-77
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    • 1990
  • 생쥐의 정자가 부정소에서 성숙하는 동안 Phosphatase 활성도의 변화를 알아보기 위하여 본 연구를 실시하였다. 효소의 활성도는 Ernst(1975)의 방법을 변용하여 측정하였으며 그 결과를 요약하면 다음과 같다. 부정소으 두부내 정자와 미부내 정자으 단백질 함량은 각각 59.1 $\pm$8.4(mg/10 9 sperm), 14.0$\pm$12.3(mg/10 9 sperm)으로 나타났다. 기본 반응액에서 나타난 두부내 정자의 효소 활성도를 100%로 할 때, 미부내 정자의 활성도는 ALPase가 29.2%, ALPase가 44.9% 그리고 ALPase가 53.8%였으며, 초음파로 정자를 분쇄하였을 경우에 ALPase는 16.5%, ALPase는 33.3% 그리고 ALPase는 38.7%였다. 따라서 미부내 정자의 phoshatase 활성도는 두부내 정자에 비하여 현저히 감소하였다. 그리고 부정소의 미부내 정자에서 K+ -dependent ALPase와 ALPase의 활성도는 두부내 정자에 비하여 유의하게 감소하였으며, 초음파로 정자를 분쇄하였을 때 $Ca^2$+ -dependent phoshatase의 활성도는 유의하게 증가하였다. 이상의 결과로 보아 생쥐의 정자가 부정소에서 성숙하는 동안 phoshatase의 활성도가 감소하는 것은 성숙과정에서 정자가 수정능력을 획득하는 데 어떤 중요한 역할을 할 것으로 사료된다.

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Kluyveromyces fragilis의 Alkaline Phosphatase 유전자의 E. coli 및 S. cerevisiae 에서의 발현 (Espression of Alkaline Phosphatase Gene from Kluyveromyces fragilis in E. coli and S. cerevisiae)

  • 박수영;황선갑;이동선;김종국;남주현;홍순덕
    • 한국미생물·생명공학회지
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    • 제23권2호
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    • pp.131-137
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    • 1995
  • The alkaline phosphatase (K-ALPase) gene of Kluyveromyces fragilis has been cloned (1) and determined its base sequences (2) previously in our laboratory. When the K-ALPase gene was expressed in Escherichia coli and Saccharomyces cerevisiae, it showed a constitutive activity in E. coli, and a derepressed activity in S. cerevisiae in phosphate-limited medium. Northem hybridization experiment was performed to elucidate the transcription level of the K-ALPase gene. Northern experiment showed that transcription level of K-ALPase gene in S. cerevisiae was higher in phosphate depletion, but it was higher in high phosphate medium than in phosphate limited medium in K. fragilis. The transcription initiation site of the K-ALPase gene was determined by primer extension analysis. It matched nucleotide position - 169 in relation to the putative trnslational start site.

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Alkaline phosphatase의 기질 특이성 (Substrate Specificity of Alkaline Phosphatase)

  • 이경태;권창호
    • 약학회지
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    • 제37권6호
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    • pp.571-576
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    • 1993
  • The substrate specificity of the purified rabbit plasma alkaline phosphatase (ALPase) was determined towards a extended range of potential substrates including relatively simple phosphate derivatives as p-NPP and indolyl phosphate, and several synthetic peptides and phosphoproteins. These results further estabilish the broad substrate specificity of these circulating enzymes. Interestingly, the plasma ALPase preferentially dephosphorylates Thr over Ser residues, as demonstrated with a series of synthetic peptides. The latter result is in contradiction to the behaviour of the tissue ALPase, which is thought to the ultimate source of plasma ALPase, and open therefore new perspectives with respective to the origin and "solubilisation" processes of these enzymes. Dephsphrylation of protein substrates by endogenous and isolated plasma ALPases indicates that ALPase probably displays protein phosphatase activity in vivo.

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신장 세포에서 Levamisole의 세포내 Alkaline Phosphatase 활성 증가 (Increase of Cellular Alkaline Phosphatase Activity by Levamisole in Kidney Cells)

  • 황준일;김종환;김주일;이경태;권창호
    • Journal of Pharmaceutical Investigation
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    • 제26권4호
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    • pp.309-314
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    • 1996
  • The purpose of this study is to explain the relationship between the pharmacological mechanism of levamisole and the cellular activity of cellular alkaline phosphatase (ALPase) in kidney cells. The results of our investigation were as follows. 1. Cellular ALPase activity in Macacus rhesus monkey kidney cells (MA 104 cells) and primary cultured rabbit kidney proximal tubular cells treated with levamisole was increased about two or three times than control. However, 50% of ALPase activity in cultured medium was inhibited by levamisole itself. 2. The proliferation of MA 104 and cultured rabbit kidney proximal tubular cells was linearly decreased in paralleled with increase of levamisole concentration $(50\;and\;500\;{mu}M)$ with MTT test. 3. In the heat stability tests, the inhibition of ALPase activity with and without levamisole at $56^{\circ}C$ in MA 104 cells showed different $IC_{50}$ values. 4. HPLC analysis of levamisole metabolites produced by cultured MA 104 cells suggested that the formation of a metabolite, that may be associated with its increase of cellular ALPase activity. Based on these results, we assumed that the increase of cellular ALPase activity by levamisole was evoked by modification of the ALPase catalytic sites.

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돼지 난포내 Alkaline Phosphatase 및 Adenosine Triphosphatase의 활성부위와 활성도 (Localization and Activity of Alkaline Phosphatase and Adenosine Triphosphatase of Ovarian Follicles in Pig)

  • 김문규;계명찬;윤현수;김종흡
    • Clinical and Experimental Reproductive Medicine
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    • 제18권2호
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    • pp.123-131
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    • 1991
  • In order to study the growth and maturation of ovarian follicle, the localization and activity of alkaline phosphatase(ALPase) and adenosine triphosphatase(ATPase) of the granulosa cells and theca layer were examined according to the follicle size, the follicle state and the ovarian cyclic phase in pig. Theca interna of the small follicles was more heavyly localized with reaction product by the activites of ALPase and ATPase than that of the large follicles. It is assumed that, as the follicles proceed to growth and maturation, antrum formation is the result of the follicular fluid accumulation by means of active transport by the activities of ALPase and ATPase in theca interna. The activities of ALPase and ATPase in atretic follicles were higher than those of normal follicles. These results imply that the mechanisms of follicle maturation and atresia are different according to the phase of ovarian cycle.

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흰쥐 초기배아 발생기간 중 수란관조직의 알카리성 Phosphatase활성도에 관한 연구 (A Study on the Activity of Alkaline Phosphatase of Rat Oviduct During Early Embryonic Development)

  • 김성례;김문규
    • Clinical and Experimental Reproductive Medicine
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    • 제18권1호
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    • pp.41-48
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    • 1991
  • The present investigation has been undertaken to elucidate the functional role of ovarian steroids on the mechanism of oviduct differentiation during early embryonic development in rat. The activity of alkaline phosphatase (ALPase) was measured in the oviduct tissue under different steroids treatment regime on day 9 pregnancy. The ALPase activity of the oviduct of pseudopregnant rat was compared with that of normal pregnant rat. The results of day 9 pregnancy rat oviduct clearly demonstrated that $17{\beta}-estradiol$ and progesterone were effective in pseudopregnant rat oviduct. In the ovary intact group the ALPase activity was similar in both of normal and pseudopregnant oviduct, but in the $17{\beta}-estradiol$ treated group the ALPase activity in normal pregnancy was significantly higher than that in pseudopregnancy. The effect of estradiol on the normal pregnant rat oviduct was apparently found on day 3 and day 9 pregnancy. This study, therefore, clearly demonstrates that $17{\beta}-estradiol$ is much potent in oviduct tissue differentiation. It is suggested that absence of $17{\beta}-estradiol$ effect on pseudopregnant rat oviduct is due to there is no embryo passing througth the oviduct.

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Saccharomyces uvarum의 Alkaline 및 Acid Phosphatase의 Isoenzyme 양상에 대하여 (Isoenzyme pattern of Aldaline and Acid Phosphatase in the Culture of Saccharomyces uvarum)

  • 이기성;최영길
    • 미생물학회지
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    • 제23권3호
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    • pp.172-176
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    • 1985
  • 본 연구는 효모세포 (Saccharomyces uvarum)의 ACPase, ALPase의 Isoenzyme type을 규명함과 아울러, iso-enzyme type중 어느 것이 constituitive 또는 repressible enzyme type인가를 조사하고, 각각의 isoenzyme활성도 비율을 살펴보아 배양조건(catabolic repression and derepression) 및 무기인산 제한에 따른 각 type의 세포내 조절기능을 분석하고자 하였다. Acid phosphatase는 또 다른 isoenzyme type이 발견되지 않았으나, alkaline phosphatase의 경우는 3가지 type이 p-NPP에 specificity를 지녔다. 세포질의 수용성단백질 분석결과 exponential phase의 세포는 주로 음전하를 띤 단백질이 높은 함량을 나타내었다. 당과 인산이 결핍된 배지에서 생육권(catabolic repression)세포에서 AL P Pase isoenzyme 중 type “B"의 활성도가 매우 높아졌으나, 완전배지에서 배양된 (catabolic derepression)세포에서는 type “B"의 활성도 감소 및 type “C"의 활성도 증가 현상을 볼 수 있었다. ALPase 중 “A" peak는 constituitive enzyme, “B" peak는 repressible enzyme, peak “C"는 L-histidinol phosphatase로 추정되었다.

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마그네슘 티타네이트 표면의 조골세포 부착도와 분화 (Osteoblast adhesion and differentiation on magnesium titanate surface)

  • 최승민;이재관;고성희;엄흥식;장범석
    • Journal of Periodontal and Implant Science
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    • 제35권4호
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    • pp.851-861
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    • 2005
  • The nature of the implant surface can directly influence cellular response, ultimately affecting the rate and quality of new bone tissue formation. The aim of this in vitro study was to investigate if human osteoblast-like cells, Saos-2, would respond differently when plated on disks of magnesium titanate and machined titanium. Magnesium titanate disks were prepared using Micro Arc Oxidation(MAO) methods. Control samples were machined commercially pure titanium disks. The cell adhesion, proliferation and differentiation were evaluated by measuring cell number, and alkaline phosphatase(ALPase) activity at 1 day and 6 day after plating on the titanium disks. Measurement of cell number and ALPase activity in Saos-2 cells at 1 day did not demonstrate any difference between machined titanium and magnesium titanate. When compared to machined titanium disks, the number of cells was reduced on the magnesium titanate disks at 6 day, while ALPase activity was more pronounced on the magnesium titanate. Enhanced differentiation of cells grown on magnesium titanate samples was indicated by decreased cell proliferation and increased ALPase activity.

가임신 흰쥐 자궁조직의 착상부위분화에 관한 연구 (A Study on the Differentiation of the Implantation Sites in the Pseudopregnant Rat Uterus)

  • 김성례
    • 한국동물학회지
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    • 제34권4호
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    • pp.479-490
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    • 1991
  • 포유류 배아의 착상기전을 규명할 목적의 일환으로 자궁내막조직의 탈락막 형성과 분화에 미치는 난소 스테로이드 호르몬의 영향과 인위적 자극등의 영향을 조사하였다. 가임신을 유도시킨 흰쥐와 정상임신군의 자궁내막조직을 착상부위와 비 착상부위로 분리하여 자궁내막조직의 분화에 지표가 되는 Alkaline phosphatase(ALPase)의 활성을 측정하였다. 가임신군에서 인위적 자극(trauma)을 가한 자궁이 자극을 받지않은(control) 자궁에 비해 Progesterone(P)만을, 또는 progesterone과 estradiol(E + P)을 동시 처리받은 실험군에서 ALPase 활성이 유의한 차이(p<0.01)로 높게 나타났다. 정상임신군에서는 착상시기인 임신 제6일군의 착상부위에서 I처리군과 P처리군의 ALPase활성이 비착상 부위에 비해 유의한 차이(P < 0.05)로 크게 나타나며, 특히 임신 제6일군에서는 착상, 비착상 부위의 P처리군의 활성이 Intact군보다 유의하게(P<0.05) 높아졌다. 정상임신 제 9일 Intact군의 ALPase활성은 임신 제3일, 6일의 Intact 군의 활성에 비하여 착상(3, 6일 :P<0.01), 비 착상부위 (3일 :P<0.05, 6일 :P<0.02)에서 다같이 유의한 차이로 높아졌다. 본 연구결과에서 배아가 분화, 발생해 감에따라 자궁내막조직 분화도 활발해지고, 착상부위 분화는 착상시기부터 현저해지며, 특히 P호르몬의 영향이 크다는 것과 배아 아닌 인위적 자극으로도 탈락막 형성 유도가 가능하다는 것을 확인할 수 있었다.

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Saccharomyces uvarum의 인삼염첨가배양에 따른 ALPase, ACPase, ATPase 활성도와 volutin과립의 함량변화 (Studies on the Changes in Activities of ALPase, ACPase, ATPase and Synthesis of Volutin Granules upon Phosphate Concentration in Saccharomyces uvarum)

  • 이기성;최영길
    • 미생물학회지
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    • 제23권2호
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    • pp.84-89
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    • 1985
  • 효모세포(Saccharomyces uvarum ATCC 9080)를 재료로, 무기인산의 농도(free, deficient, excess)를 달리 적용하여 배양하였을 경우, 세포내 무가폴리인산의 축적량, ortho-P, nucleotidic labile-P의 함량 및 ALPase, ACP Pase, ATPase의 활성도 변화을 관련지어. catabolic repression시킨 세포와 catabolic derepression시킨 세포의 인산대사 조절에 대한 무기인산 농도의 영향를 살피고자 하였다. 무기인산을 결핍시킨 배지에서 배양된 세포(PO)는 배지에 당을 첨가하면 분열속도는 늦지만 세포분열이 가능하고, 무기폴리인산 축적이 감소되었다. 인산의 첨가 농도에 관계없이 무기폴리인산 각형의 함량변화(전환)는 있었지만 총무기폴리인산의 함량은 거의 유사하였다. 정상배양세포에 비하여, 당은 공급되었지만 인산은 결핍시킨 배지에서 배양한 세포에서 ALPase, ACPase, ATPase의 활성도가 지속적으로 높았다. 무기인산 결핍배지에서 배양한 세포내에 존재하는 무기폴리인산계의 기능이 특히 당과 무기인산 결핍(catabolic repression)시기에 무기인산 공여체, 에너지원 및 regulator로 ATP/ADP system의 기능을 보상작용하는 것으로 나타났다.

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