• Title/Summary/Keyword: ALPase

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

  • 김문규;윤현수;김종흡;김성례
    • The Korean Journal of Zoology
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    • v.33 no.1
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    • pp.70-77
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    • 1990
  • The change of phosphatase activities of the epididymal spermatozoa has been examined during epididymal maturation in mouse. The quantitative analysis of phQsphatase activities have been carried out using the method modified by Emst(1975). The results of experiment were summarized as the followings. Total protein of the caput epididyrnal spermatozoa(CPS) was measured as 59.1 $\pm$8.4(mg/10 9 spermatozoa), and that of the cauda epididymal spermatozoa(CDS) was 14.0$\pm$12.3(mg/10 9 spermatozoa). When phosphatase activities of the CDS in basic reaction medium were 29.2% in alkaline phosphatase, 44.9% in ATPse and 53.8% in acid phosphatase. The activities were eminently decreased in all CDS in contrast to those of CPS. The alkaline phosphatase and ATPase activities of K+ -dependent were decreased in CDS when compared with caput epididymal spermatozoa, and alkaline phosphatase, ATPase and acid phosphatase activities of $Ca^2$+ -dependent were increased in homogenized spermatozoa when compared with intact spermatozoa. From these results, it may be concluded that the decrease of phosphatases activities in spermatozoa during epididymal maturation may play some significant roles in acquiring fertilizing capability.

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

  • Park, Soo-Young;Hwang, Seon-Kap;Lee, Dong-Sun;Kim, Jong-Guk;Nam, Joo-Hyun;Hong, Soon-Duck
    • Microbiology and Biotechnology Letters
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    • v.23 no.2
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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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Substrate Specificity of Alkaline Phosphatase (Alkaline phosphatase의 기질 특이성)

  • ;;E. Waelkens;W. Merlevede
    • YAKHAK HOEJI
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    • v.37 no.6
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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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Increase of Cellular Alkaline Phosphatase Activity by Levamisole in Kidney Cells (신장 세포에서 Levamisole의 세포내 Alkaline Phosphatase 활성 증가)

  • Hwang, Joon-Il;Kim, Jong-Hwan;Kim, Joo-Il;Lee, Kyung-Tae;Kwon, Chang-Hoo
    • Journal of Pharmaceutical Investigation
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    • v.26 no.4
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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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Localization and Activity of Alkaline Phosphatase and Adenosine Triphosphatase of Ovarian Follicles in Pig (돼지 난포내 Alkaline Phosphatase 및 Adenosine Triphosphatase의 활성부위와 활성도)

  • Kim, Moon-Kyoo;Gye, Myung-Chan;Yoon, Hyun-Soo;Kim, Jong-Heup
    • Clinical and Experimental Reproductive Medicine
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    • v.18 no.2
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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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A Study on the Activity of Alkaline Phosphatase of Rat Oviduct During Early Embryonic Development (흰쥐 초기배아 발생기간 중 수란관조직의 알카리성 Phosphatase활성도에 관한 연구)

  • Kim, Sung-Rye;Kim, Moon-Kyoo
    • Clinical and Experimental Reproductive Medicine
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    • v.18 no.1
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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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Isoenzyme pattern of Aldaline and Acid Phosphatase in the Culture of Saccharomyces uvarum (Saccharomyces uvarum의 Alkaline 및 Acid Phosphatase의 Isoenzyme 양상에 대하여)

  • 이기성;최영길
    • Korean Journal of Microbiology
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    • v.23 no.3
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    • pp.172-176
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    • 1985
  • The present study was designed to investigate isoenzyme (ACPase, ALPase) pattern and its refulatory function between catabolically repressed and derepressed states in yeast, Saccharomyces uvarum. As the results, no other isoenzyme was detectable in acid phosphatase, but there were three isoenzyme types in aldaline phosphatase. Type "B" isoenzyme among alkaline phosphatases in catabolically repressed cell was derepressed, but in normally cultivated cell, type "C" isoenzyme was derepressed while type "B" activity was lowered. Type "B" isoenzyme could be postulated as repressible enzyme, type "A" as constityityve enzyme and type "C" as L-histidinol phosphatase, respectively, Also, it could be shown that type "B" ALPase, repressible enzyme, compensated for phosphate group supplier under catabolically repressed states. Protein profile in cytoplasmic soluble fraction of exponential phase cell was characterized by negative charged protein.

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

  • Choi, Seung-Min;Lee, Jae-Kwan;Ko, Sung-Hee;Um, Heung-Sik;Chang, Beom-Seok
    • Journal of Periodontal and Implant Science
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    • v.35 no.4
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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.

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

  • 김성례
    • The Korean Journal of Zoology
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    • v.34 no.4
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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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Studies on the Changes in Activities of ALPase, ACPase, ATPase and Synthesis of Volutin Granules upon Phosphate Concentration in Saccharomyces uvarum (Saccharomyces uvarum의 인삼염첨가배양에 따른 ALPase, ACPase, ATPase 활성도와 volutin과립의 함량변화)

  • 이기성;최영길
    • Korean Journal of Microbiology
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    • v.23 no.2
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    • pp.84-89
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    • 1985
  • The effect of exogenous phosphate supply on the regulation of phosphate metabolism was investegated during catabolic repression and catabolic derepression in yeast (Saccharomyces uvarum). As the results, when sugar was supplimented in cells cultivated under phosphate free, the growith rate was low but it was capable of cell division. Polyphosphate "B" was accumulated highly in proportion to amount of phosphate added to the medium. Without regard to phosphate supply of the medium, the total amount of polyphospgate was almost similar, although each polyphosphate was turned over. Activities of all phosphatases remained continuousoy high in the cells cultivated in the phosphate free medium. Especially under catabolic repression, the function of polyphosphate system was shown to compensate the ATP/ADP system as phosphate donor, energy source and regulator.

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