• Title/Summary/Keyword: Acid Phosphatase

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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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Identification and Characterization of the Acid Phosphatase HppA in Helicobacter pylori

  • Ki, Mi-Ran;Yun, Soon-Kyu;Choi, Kyung-Min;Hwang, Se-Young
    • Journal of Microbiology and Biotechnology
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    • v.21 no.5
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    • pp.483-493
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    • 2011
  • An acid phosphatase (HppA) activated by $NH_4Cl$ was purified 192- and 34-fold from the periplasmic and membrane fractions of Helicobacter pylori, respectively. SDS-polyacrylamide gel electrophoresis revealed that HppA from the latter appears to be several kilodaltons larger in molecular mass than from the former by about 24 kDa. Under acidic conditions (pH${\leq}$4.5), the enzyme activity was entirely dependent on the presence of certain mono- and/or divalent metal cations (e.g., $K^+$,$ NH_4{^+}$, and/or $Ni^{2+}$). In particular, $Ni^{2+}$ appeared to lower the enzyme's $K_m$ for the substrates, without changing $V_{max}$. The purified enzyme showed differential specificity against nucleotide substrates with pH; for example, the enzyme hydrolyzed adenosine nucleotides more rapidly at pH 5.5 than at pH 6.0, and vice versa for CTP or TTP. Analyses of the enzyme's N-terminal sequence and of an $HppA^-$ H. pylori mutant revealed that the purified enzyme is identical to rHppA, a cloned H. pylori class C acid phosphatase, and shown to be the sole bacterial 5'-nucleotidase uniquely activated by $NH_4Cl$. In contrast to wild type, $HppA^-$ H. pylori cells grew more slowly. Strikingly, they imported $Mg^{2+}$ at a markedly lowered rate, but assimilated urea rapidly, with a subsequent increase in extracellular pH. Moreover, mutant cells were much more sensitive to extracellular potassium ions, as well as to metronidazole, omeprazole, or thiophenol, with considerably lowered MIC values, than wild-type cells. From these data, we suggest that the role of the acid phosphatase HppA in H. pylori may extend beyond 5'-nucleotidase function to include cation-flux as well as pH regulation on the cell envelope.

Effects on Ultrastructural Changes and Several Phosphatase Activities by Actinomycin D in Migrating Primordial Germ Cells of Developing Rat (발생중인 흰쥐 원생식세포의 이동과 미세구조 및 몇몇 Phosphatase의 활성에 미치는 Actinomycin D의 영향)

  • Choi, C.K.
    • Applied Microscopy
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    • v.15 no.1
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    • pp.1-12
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    • 1985
  • In this study, the pathway and date of migrating Primordial germ cells (PGCs) were observed light microscopically and ultrastructural changes of them during migration were observed by electron microscopic examination. For these purpose, alkaline phosphatase reactions were used for identifying the PGCs and acid phosphatase reactions were used for observing their degenerating activities. Also, effects of actinomycin D on the migration of PGCs were examined. According to these results, at the 9th gestation day, PGCs were observed in the endodermal cells of yolk sac, at the 11th gestation day, they were seen in the hindgut and then entered into the dorsal mesentery by the 13th gestation day. At the 14th gestation day, they were located in the genital ridges. When PGCs were located in the hindgut and genital ridges, the positive reactions of alkaline phosphatase were dominated, but acid phosphatase reactions were limited in all stage except they were in dorsal mesentery. However, these reactions were lessened in case of actinomycin D treatment. By electron microscopic examination, PGCs had pseudopodia, tail process, trailing cytoplasm and nuage as the ultrastructural characteristics. In addition, these morphological features were damaged by actinomycin D treatment.

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Celluomonas sp. AP-7이 생산하는 Ascorbic Acid Phosphorylating Enzyme의 정제 및 특성

  • 이상협;최현일;방원기
    • Microbiology and Biotechnology Letters
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    • v.25 no.3
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    • pp.271-276
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    • 1997
  • An ascorbic acid phosphorylating enzyme, which catalyzes the formation of ascorbic acid-2-phosphate from ascorbic acid and pyrophosphate, was purified 32.7-folds to homogeneity from a cell-free extract of Cellulomonas sp. AP-7. The combination of DEAE- Sephacel ion exchange chromatography and Sephacryl S-200 get filtration was used for their purification. The molecular weight of the native protein was estimated to be 96.lkDa on high performance gel filtration chromatography. The SDS-PAGE analysis indicated that the protein consisted of four identical subunits of 24.6 kDa. The purified enzyme showed the optimal tempeature of 40$\circ$C and optimal pH of 4.5. The Km for ascorbic acid and pyrophosphate were 119 mM and 11.9 mM, respectively. The addition of 5,5'-dithiobis-(2-nitrobenzoic acid) into the reaction mixture resulted in the reduction of the enzyme activity at 51%. The enzyme also had a phosphatase activity at weakly acidic pH and the Km for ascorbic acid-2-phosphate in phosphatase activity was 7.9 mM.

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Purification and Characterization of ATPase and Phosphatase of Light Membrane Vesicles Isolated from Cucurbita pepo (Cucurbita pepo에서 분리한 Light Membrane Vesicle의 ATPase와 Phosphatase의 정제 및 특성)

  • 오승은
    • Journal of Plant Biology
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    • v.33 no.4
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    • pp.325-332
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    • 1990
  • Light membrane vesicles were isolated from the zucchini hypocotyl by floatation on ficoll density gradients and the proteins were solubilized with Triton X100. Three ATP-hydrolyzing enzymes were partially purified by ion-exchange and gel filtration chromatography and isoelectric focusing. There are plasma membrane-type ATPase whose activity was inhibited by vanadate but not by nitrate, tonoplast-type ATPase which was sensitive to nitrate but insensitive to vanadate and one having a phosphatase activity with a pI value different from that of an acid phosphatase. A fraction was obtained after DEAE-ion-exchange chromatography crossreacting with polyclonal antibodies against Ca2+ -ATPase from human erythrocytes.

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A Cytochemical Study on the Acid Phosphatase of Neurons of Developing Chick Embryo Brain (계배 뇌 발생에 있어서 신경세포의 Acid Phosphatase 변화에 관한 효소화학적 연구)

  • Koh, Ki-Seok;Shin, Chu-Og;Yoo, Chang-Kyu;Choe, Rim-Soon
    • Applied Microscopy
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    • v.18 no.2
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    • pp.119-131
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    • 1988
  • The purpose of this study was to investigate the differentiation and degeneration of neurons in developing chick embryo. The activity of acid phosphatase(ACP) was measured and cytochemical study of ACP and ultrastructural changes were observed in prosencephalon, mesencephalon and rhombencephalon from day 4 to day 19 of incubation. As a result, the activity of ACP of all brain region was tend to increase from day 4 to day 19. On day 13, activities of ACP of mesencephalon and rhombencephalon were increased greatly and activity of ACP was decreased each region on day 17. On electron microscopic examination, the reaction product of ACP were localized at GERL complex, lysosome, Golgi body and vacuoles of neurons. Morphologically, disrupted nuclear envelope, mitochondrial destruction, vacuolization and ribosomal crystalization were observed.

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Evaluation and Selection of Potential Parents Based on Selection Indices and Isozyme Variability in Silkworm, Bombyx mori, L.

  • Moorthy S.M.;Das S.K.;Rao, P.R.T.;Urs S. Rao,;Sarkar A.
    • International Journal of Industrial Entomology and Biomaterials
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    • v.14 no.1
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    • pp.1-7
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    • 2007
  • In order to find out the appropriate parents for the breeding programme, twelve bivoltine and three multivoltine silkworm breeds were evaluated on the basis of multivariate selection index and isozyme analysis. Of which, four [CSR2, D6 (P), SK3, SK4] bivoltine and two multivoltine (Nistari, Cambodge) breeds were selected and breeding initiated to develop higher survival bivoltine silkworm breed suitable for tropical conditions. Among two isozyme (Esterase and acid phosphatase) analyzed, only esterase exhibited polymorphism among the bivoltine breeds. No polymorphism was observed among multivoltine in respect of esterase as well as acid phosphatase.

THE EFFECTS OF MECHANICAL STRESS ON CULTURED BONE CELL POPULATIONS (Mechanical stress가 골조직세포군에 미치는 영향)

  • Kim, Sang-Tae;Cha, Kyung-Suk
    • The korean journal of orthodontics
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    • v.24 no.1 s.44
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    • pp.105-114
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    • 1994
  • The movement of teeth during orthodontic treatment requires bone remodeling process of bone formation and bone resolution. To find out the changes occuring in the cell itself, mechanical stress was applied to the cell populations involved in the bone metabolism. Bone tissue cell populations were isolated from fetal rat calvaria and divided into OC and OB groups. Following results were obtained from measuring the changes in acid & alkaline phosphatease activity, cyclic AMP and $PGE_2$ production in time lapse after the application of mechanical stress. 1. In case of the marker enzyme of specific bone tissue cell, acid phosphatase activity was high in OC group and alkaline phosphatase activity was high in OB group. 2. After the mechanical stress was applied, acid phosphatase activity was decreased in both OC and OB groups and alkaline phosphatase activity was increase in OB group. 3. When the mechanical stress was applied for 15, 30 and 60 minutes, the production of $PGE_2$ increased in both OC and OB groups, as the time span increased. 4. When the mechanical stress was applied for 20 and 40 minutes, the production of $PGE_2$ increased in both OC and OB groups, as the time span increased.

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A Comparative Study on the Effects of Polygoni Radix and Cynanchi Radix on Rat Livers Intoxicated with Carbon Tetrachloride (백서(白鼠) 간조직(肝組織)에 미치는 적하수오(赤何首烏)와 백하수오(白何首烏)의 효능에 관한 비교(比較) 연구(硏究))

  • Shin, Min-Kyo
    • Korean Journal of Pharmacognosy
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    • v.16 no.2
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    • pp.81-92
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    • 1985
  • The Polygoni Radix and Cynanchi Radix have been used to potentiate the liver functions in clinic of Oriental Medicine. The water extracts of Polygoni Radix and Cynanchi Radix were administered orally to rats intoxicated with carbon tetrachloride and then this experiment have been performed by observing liver fatty degeneration and activities of enzymes such as cytochrome oxidase (CYO), adenosine triphosphatase (ATP), acid phosphatase (ACP), lactate dehydrogenase (LDH) and alkaline phosphatase (ALP). By oral administration of water extracts of the radices between 1 and 10 days, the following results were obtained. 1. The group given Cynanchi Radix extract showed recovery of the fat liver in 4 days, whereas that given Polygoni Radix extract did the recovery in 8 days. 2. In cytochrome oxidase activity, the group given Cynanchi extract showed normal activity in 6 days, whereas that given Polygoni Radix extract did the activity in 8 days. 3. In adenosine triphosphatase activity, the groups given Cynanchi Radix and Polygoni Radix extracts showed normal activities in 2 and 8 days, respectively. 4. In acid phosphatase activity, the groups given Cynanchi Radix and Polygoni Radix extracts showed recovery of the activities in 2 and 4 days, respectively. 5. In lactate dehydrogenase activity, the group given Cynanchi Radix and Polygoni Radix extracts showed recovery of the activities in 6 and 10 days, respectively. 6. In alkaline phosphatase activity, the group given Cynanchi Radix extract showed normal activity in 2 days, whereas that given Polygoni Radix extract showed slight recovery between 4 and 6 days followed by decrease of the activity in 8 and 10 days. From the above-mentioned results, it was found that both of the water extracts of Polygoni and Cynanchi Radix possessed the recovery action of liver function as intoxicated with carbon tetrachloride in rats. It is also noted that the extract of Cynanchi Radix showed more potent activity than that of Polygoni Radix.

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