• Title/Summary/Keyword: ALPase activity

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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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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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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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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.

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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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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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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The biologic effects of magnoliae cortex extract and safflower seed (Carthamus tinctorius $Linn{\acute{e}}$) extract mixture on PDL cells and osteoblasts (후박 및 홍화종자 추출혼합물이 치주인대세포 및 골아세포의 활성도 및 백서의 두개골재생에 미치는 영향)

  • Shin, Seung-Yun;Lee, Yong-Moo;Ku, Young;Bae, Ki-Hwan;Chung, Chong-Pyoung
    • Journal of Periodontal and Implant Science
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    • v.28 no.4
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    • pp.545-559
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    • 1998
  • Magnoliae cortex has been used as a drug for treatment of fractures in Chinese medicine and safflower(Carthamus tinctorius $Linn{\acute{e}}$) has been traditionally used for treatment of blood stasis. The purpose of present study was to examine the biologic effects of magnoliae cortex extract and safflower extract mixture(MSM) on human periodontal ligament cells and fetal rat calvarial osteoblasts and on healing of rat calvarial defects. The ethanolic extracts of magnoliae cortex(MCE), safflower seed(SSE), Zea May L(ZML) were prepared as positive control group. MSM mixed to the ratios of 1 : 1, 1 : 2, 1 : 5 and 1 : 10 were used as test group. The effects of each agents on the growth and survival, ALPase activity, cell proliferation and tissue regenerative effect of each extracts was evaluated by histomorphometric measuring of newly formed bone on the 8 mm defect in rat calvaria after oral administration of 2 ratio groups(1 : 5 and 1 : 10) at 3 different doses (0.1, 0.25 and 0.5g/kg per day). MSM stimulated the growth and survival rate of osteoblasts and PDL cells more than any other agents. The growth and survival rate were increased as the proportion of safflower seed extract was increased. MCE, SSE, ZML stimulated the ALPase activity of osteoblast and PDL cell in comparison to the negative control group. But all groups of MSM regardless of ratio of safflower seed extract stimulated the ALPase activity than any other agent. The ALPase activity was also increased as the proportion of safflower seed extract was increased. Although MCE, SSE, ZML stimulated the proliferation of osteoblasts. 1 : 5 and 1 : 10 ratio MSM showed significant increase in stimulation of proliferation of osteoblasts. No agent significantly increased proliferation of PDL cells. Significant new bone formation were seen where 1 : 5 ratio, 0.5g/kg group and 1 : 10 ratio, 0.25, 0.5g/kg groups were used. These results show that magnoliae cortex extract and safflower seed extract mixture can potentially increase bone regeneration ability.

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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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Changes of Digestive Enzyme Activity in Slime Flounder, Micromus achne Larvae with Growth and Development (찰가자미, Micromus achne 자어의 성장과 발달에 따른 소화효소 활성의 변화)

  • 박상언;임한규;한현섭;이종하;임영수;이종관;이상민
    • Journal of Aquaculture
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    • v.16 no.4
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    • pp.233-239
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    • 2003
  • Growth and activities of digestive enzymes in slime flounder (Microstomus achne) larvae were measured from hatching to near the end of larval development (day 58). Larvae reared under starved and fed conditions and the changes of acid phosphatase (ACPase) specific activity, alkaline phosphatase (ALPase) specific activity, trypsin-like enzyme activity and pepsin-like enzyme activity were described with growth and developmental stage of larvae. Total length of the starved larvae was gradually increased for 7 days post hatching and then almost unchanged. Total length of the fed larvae ranged from 5.13$\pm$0.178 mm at the day of hatching to 13.43$\pm$1.395 mm at 58 days after hatching. In starved group, dry body weight decreased from 0.l0$\pm$0.020 mg at the day of hatching to 0.05$\pm$0.012 mg at 12 days after hatching. Dry body weight of fed larvae decreased during the prelarva stage like starved group and then gradually increased. ACPase and ALPase specific activity in the starved larvae increased until all larvae died, however those activities in the fed larvae increased until 20 days and then decreased until 58 days after hatching, with no significant difference between groups. Trypsin-like enzyme activity in the starved larvae was unchanged until 3 days and then was the highest on 5 days after hatching, but not detected after completion of yolksac absorption. Those of fed larvae decreased until 3 days and sharply increased until completion of yolksac absorption. The highest trypsin-like enzyme activity in the fed group was observed at 20 days after hatching. Trypsin-like enzyme activity in the fed larvae was significantly higher than that in the starved larvae from 8 days after hatching. Pepsin-like enzyme activity was increased in 5 days after hatching in both groups. There was significant difference at 8 and 10 days after hatching between both groups. Based on above results, digestive enzyme activities were correspondingly changed to a growth and morphological transformation. Trypsin-like enzyme and pepsin-like enzyme activities are able to be a useful indices for health and growth status in larval slime flounder, because there was significant difference in digestive enzyme activities with developmental stages, growth or feed supply.