• Title/Summary/Keyword: phosphatase

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Lysosomal Acid Phosphatase in Regenerating Salamander Limbs Studied with Monoclonal Antibodies (리소솜 Acid Phosphatase에 대한 단일 항체를 이용한 도롱뇽 다리 재생 연구)

  • 주봉건;박상렬;최의열;김원선
    • The Korean Journal of Zoology
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    • v.39 no.4
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    • pp.426-436
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    • 1996
  • In previous studies, we have shown that lysosomal add phosphatase (LAP) activity increases at the dedifferentiation stage in the regenerating larval limbs of salamander, Hynobius leechii. Monoclonal antibodies against LAP were generated to determIne the spatial and temporal distribution of the protein In the regenerates.A total of 22 monoclonal antihodies recognizIng different epftopes of the protein were obtained, of which five strongly stained the regenerating limb by imunohistochemistry. in LAP immunohistochemical examination, LAP showed distribution coincident with the state of dedifferentiation, both spatially and temporally, in the limb regenerates. When unfractioned protein of regenerating salamander limbs were separated by gel electrophoresis and immunoblotted, the antibodies recognized a single protein band of 53 kl)a, which comigrates with a monomerlc subunit of IAR Using the anti-IAP antibodIes as probe, we investigated the cross-reactivities of LAPs from other sources. The immunoreadive bands on Western blots appeared to be the same In molecular mass-53 kl)a in axoloti and Xenopus, but no protein band was detected in mouse, Drosophila, or C. elegans.These results show that the antibodies generated in this study spedfically recognize Hynoblus leeclili IAp and that IAPs may be highiy conserved among amphibians. Furthermore, the distdbution of the protein is consistent with a role for LAP in the dedifferentiation process of limb regeneration.

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The effect of low level laser irradiation on proliferation and alkaline phosphatase activity of human gingival fibroblast in vitro (저출력 레이저 조사가 치은섬유아세포의 증식과 염기성 인산분해효소의 활성에 미치는 영향)

  • Park, Byung-Ki;Lim, Kee-Jung;Kim, Byung-Ock;Han, Kyung-Yoon
    • Journal of Periodontal and Implant Science
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    • v.26 no.3
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    • pp.715-724
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    • 1996
  • This study was performed to identify the proliferation and to measure the alteration of alkaline phosphatase activity in human gingival fibroblasts cultured. For the present study, the authors cultured the human gingival fibroblasts oriented from the sound interdental gingiva, and used third passage. It was used methyl $[^3H]$ Thymidine to identify the proliferation in human gingival fibroblasts and used 410nm of the spectrophotometer to measure the alteration of the alkaline phosphatase activity in human gingival fibroblasts. The results were as follows: 1. There was a statistically significant increase in the proliferation of gingival fibroblasts following low level laser irradiation at 24 hour(p<0.05). 2. There was a statistically significant increase in activity of alkaline phosphatase compared to control group at 5-day laser irradiation after in laser irradiation groups(p<0.05). And there was a statistically significant increase in activity of alkaline phosphatase compared to control group at 7-day laser irradiation after in the I-minute laser irradiation group(p<0.05), but there was a statistically significant decrease in activity of alkaline phosphatase compared to 1minute laser irradiation group at 7-day laser irradiation in the 2-minute laser irradiation group after(p<0.05). The results, within the limits of the present experiments, suggest that, the low level laser irradiation accelerates the proliferation of gingival fibroblasts and alters the alkaline phosphatase activity until the restricted period.

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The Effects of Phosphate Starvation on the Activities of Acid and Alkaline Phosphatase, Fructose-1,6-bisphosphatase, Sucrose-phosphate Synthase and Nitrate Reductase in Melon (Cucumis melo L.) Seedlings

  • Kang, Sang-Jae;Lee, Chang-Hee;Park, Man
    • Korean Journal of Soil Science and Fertilizer
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    • v.49 no.1
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    • pp.44-52
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    • 2016
  • Plants response to phosphate starvation include the changes of activity of some enzymes, such as phosphatases, fructose-1,6-bisphosphatase, sucrose-phosphate synthase and nitrate reductase. In this study, to determine the effects of phosphate starvation on the change of activities of acid and alkaline phosphatase, fructose-1,6-bisphosphatase, sucrose-phosphate synthase, and nitrate reductase were studied in melon seedlings (Cucumis melo L.). The content of the protein and chlorophyll tended to relatively reduced in melon seedlings subjected to phosphate starvation. Acid phosphatase activity in first and second leaves of melon seedlings was relatively higher than that of third and fourth leaves of seedlings in 14 days after phosphate starvation treatment, respectively. Active native-PAGE band patterns of acid phosphatase in melon leaves showed similar to activities of acid phosphatase, whereas alkaline phosphatase activity was different from the change in the activity of acid phosphatase. Inorganic phosphate content in melon seedlings leaves was constant. The changes of Fructose-1,6-bisphosphatase and sucrose phosphate synthase activities showed similar patterns in melon seedlings leaves, and between these enzymes activities and phosphate nutrition negatively related. Fructose-1,6- bisphosphatase and sucrose phosphate synthase activities showed significant difference in second and fourth leaves, but nitrate reductase showed significant difference in first and second leaves in 14days after phosphate starvation treatment. We concluded that phosphate nutrition could affect the distribution of phosphate, carbon and nitrogen in melon seedlings.

Zinc Deficiency Decreased Alkaline Phosphatase Expression and Bone Matrix Ca Deposits in Osteoblast-like MC3T3-E1 Cells

  • Cho Young-Eon;Lomeda Ria-Ann R.;Kim Yang-Ha;Ryu Sang-Hoon;Choi Je-Yong;Kim Hyo-Jin;Beattie John H.;Kwun In-Sook
    • Nutritional Sciences
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    • v.8 no.4
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    • pp.242-249
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    • 2005
  • It is well established that zinc plays an important role in bone metabolism and mineralization. The role of zinc in bone formation is well documented in animal models, but not much reported in cell models. In the present study, we evaluated zinc deficiency effects on osteoblastic cell proliferation, alkaline phosphatase activity and expression, and extracellular matrix bone nodule formation and bone-related gene expression in osteoblastic MC3T3-E1 cells. To deplete cellular zinc, chelexed-FBS and interpermeable zinc chelator TPEN were used. MC3T3-E1 cells were cultured in zinc concentration-dependent (0-15 ${\mu}M\;ZnCl_2$) and time-dependent (0-20 days) manners. MC3T3-E1 cell proliferation by MTT assay was increased as medium zinc level increased (p<0.05). Cellular Ca level and alkaline phosphatase activity were increased as medium zinc level increased (p<0.05). Alkaline phosphatase expression, a marker of commitment to the osteoblast lineage, measured by alkaline phosphatase staining was increased as medium zinc level increased. Extracellular calcium deposits measured by von Kossa staining for nodule formation also appeared higher in Zn+(15 ${\mu}M\;ZnCl_2$) than in Zn-(0 ${\mu}M\;ZnCl_2$). Bone formation marker genes, alkaline phosphatase and osteocalcin, were also expressed higher in Zn+ than in Zn-. The current work supports the beneficial effect of zinc on bone mineralization and bone-related gene expression. The results also promote further study as to the molecular mechanism of zinc deficiency for bone formation and thus facilitate to design preventive strategies for zinc-deficient bone diseases.

General Enzymatic Properties of Human Histidine Acid Phosphatase-Phytase (히스티딘 에시드 포스파테이즈(Histidine Acid Phosphatase) 계열 인간 파이테이즈(Phytase)의 일반적 특성규명)

  • Cho, Jaie-Soon
    • Journal of Animal Science and Technology
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    • v.51 no.2
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    • pp.177-182
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    • 2009
  • The glycosylated human MINPP (multiple inositol polyphosphate phosphatase), which was recombinantly over-expressed by using industrial host, Pichia pastoris, showed the phytase activity against phytate ($InsP_6$) and the enzyme activity of the unglycosylated counterpart was decreased to 30%. The optimal phytase activity occurred at pH 7.4. The human MINPP showed high substrate specificity for $InsP_6$ with little activity on other organic phosphate conjugates such as para-nitrophenylphosphate (pNPP), ATP, and ribose-1-phosphate (R-1-P). The phosphatase activity against 2,3-bisphosphoglycerate (2,3-BPG) by human MINPP was increased to 1.2-fold in the presence of stimulator, 1 mM 2-phosphoglycolate (2-PG) but the phytase activity against $InsP_6$ was not affected by addition of 1 mM 2-PG. The phosphatase activity against 2,3-BPG by human MINPP was not increased in the presence of 2 mM $Mg^{2+}$ or 100 mM $Cl^-$.

Purification and Crystallization of the Recombinant Catalytic Subunit of Pyruvate Dehydrogenase Phosphatase (Pyruvate Dehydrogenase Phosphatase의 Catalytic Subunit의 분리정제 및 결정화)

  • Kim, Young-Mi
    • Journal of Food Hygiene and Safety
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    • v.18 no.3
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    • pp.146-152
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    • 2003
  • Pyruvate dehydrogenase phosphatase (PDP) is a mitochondrial protein serine/threonine phosphatase that catalyzes the dephosphorylation and concomitant reactivation of the pyruvate dehydrogenase component of the pyruvate dehydrogenase complex (PDC). PDP consists of a catalytic subunit (PDPc, Mr 52,600) and regulatory subunit (PDPr, Mr 95,600). In the presence of $Ca^{2+}$, PDPc binds to the dihydrolipoamide acetyltransferase (E2) component of the pyruvate dehydrogenase complex in proximity to its substrate, the phosphorylated E1 component, thereby increasing the rate of dephosphorylation. PDPc possesses and intrinsic $Ca^{2+}$ binding site and a second $Ca^{2+}$ site is generated in the presence of E2. Using the unique interaction, highly pure PDPc was produced by the GSH-Sepharose-GST-L2 matrix with a specific activity of approx. 1000 U/mg and a yield of about 80%.

Studies on the application of bone alkaline phosphatase and percentage of BALP values to total alkaline phosphatase as a marker of bone formation (골혈성 지표로써 bone alkaline phosphatase(BALP)와 TALP치에 대한 BALP치 비율(B/T)의 응용)

  • Kim, Nam-soo;Choi, In-hyuk
    • Korean Journal of Veterinary Research
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    • v.39 no.6
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    • pp.1197-1209
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    • 1999
  • Twenty-one dogs(male 11 heads, female 10 heads) which were about 16 months ($16.3{\pm}3.5$) old and 10kg($10.1{\pm}2.0$) body weight, were allotted randomly into four groups as follows. Group I consisted of five dogs whose muscles were operated for sham muscle injuries. Group II consisted of seven dogs treated for cystic duct obstruction. Group III consisted of five dogs treated for the union fracture model. Group IV consisted of five dogs treated for the non-union fracture model. Radiographical and histological observations were carried out to determine bone alkaline phosphatase(BALP) and total alkaline phosphatase(TALP) values of each group for 20 weeks after the treatments with the condition of new bone formation. And also the applicability of percentage of BALP values to TALP (B/T) was studied after BALP was compared respectively with TALP. The level of TALP was increased without any relation to bone formation in group II, and all levels of BALP and B/T were increased in group III. The mean of B/T was high in statistical significance, due to varied levels of B/T and BALP. The changes of rates of B/T were significantly increased only in the case of the active new bone formation in group III, union fracture model. It was recognized that the mean values of B/T were statistical significant of the high applicability of the B/T ratio as an index of bone formation.

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Ultrastructural Localization of Acid Phosphatase in Entamoeba histolytica and Entamoeba gingivalis (이질아메바 (Entamoeba histolytica)와 치은아메바 (Entamoeba gingivalis)의 Acid Phosphatase 활성에 관한 세포화학적 연구)

  • Cho, Kee-Mok;Cha, Hai-Young;Soh, Chin-Thack
    • Applied Microscopy
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    • v.3 no.1
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    • pp.1-16
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    • 1973
  • A combined cytochemical and electron microscopic study was carried out for the demonstration of acid phosphatase activities in trophozoites of E. histolytica. and E. gingivalis. E. histolytica(YS-27) strain was isolated from liver abscess of 72-year-old man in September 1969, and E. gingivalis (YS-215) strain was collected from gingival crevice of 41-year-old man in January 1972. The amoeba strains were maintained by subculture on diphasic medium, and used throughout the study. The results are summarized as follows; 1. In E. histolytica, the reaction products were distributed evenly over the entire surface of plasma membrane, whereas E. gingivalis showed no activity of acid phosphatase on the plasma membrane, except in the portion of the uroid-like structure. 2. In the cytoplasm, various reaction precipitates were observed in vacuoles of both amoebae; vacuole limiting membrane, vacuole membrane and its contents and lysosome-like structure. Strong enzyme active contents but membrane reaction negative vacuoles were conspicuous in E. gingivalis. Endoplasmic reticulum showed a moderate activity. 3. Granule-like acid phosphatase reaction product was demonstrated in the nucleoplasm of E. gingivalis, but it was negative in E. histolytica.

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Emerging Roles of CTD Phosphatases (CTD 탈 인산화 효소의 기능과 역할)

  • Kim, Youngjun
    • Journal of Life Science
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    • v.27 no.3
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    • pp.370-381
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    • 2017
  • Protein dephosphorylation is important for cellular regulation, which is catalyzed by protein phosphatases. Among protein phosphatases, carboxy-terminal domain (CTD) phosphatases are recently emerging and new functional roles of them have been revealed. There are 7 CTD phosphatases in human genome, which are composed of CTD phosphatase 1 (CTDP1), CTD small phosphatase 1 (CTDSP1), CTD small phosphatase 2 (CTDSP2), CTD small phosphatase-like (CTDSPL), CTD small phosphatase-like 2 (CTDSPL2), CTD nuclear envelope phosphatase (CTDNEP1), and ubiquitin-like domain containing CTD phosphatase 1 (UBLCP1). CTDP1 dephosphorylates the second phosphor-serine of CTD of RNA polymerase II (RNAPII), while CTDSP1, STDSP2, and CTDSPL dephosphorylate the fifth phosphor-serine of CTD of RNAPII. In addition, CTDSP1 dephosphorylates new substrates such as mothers against decapentaplegic homologs (SMADs), cell division cycle-associated protein 3 (CDCA3), Twist1, tumor-suppressor protein promyelocytic leukemia (PML), and c-Myc. CTDP1 is related to RNA polymerase II complex recycling, mitosis regulation and cancer cell growth. CTDSP1, CTDSP2 and CTDSPL are related to transcription factor recruitment, tumor suppressor function and stem cell differentiation. CTDNEP1 dephosphorylates LIPIN1 and is related to neural tube formation and nuclear envelope formation. CTDSPL2 is related to hematopoietic stem cell differentiation. UBLCP1 dephosphorylates 26S proteasome and is related to nuclear proteasome regulation. In conclusion, noble roles of CTD phosphatases are emerging through recent researches and this review is intended to summarize emerging roles of CTD phosphatases.

The effects of continuous and intermittent compressive pressure on alkaline phosphatase activity of Periodontal Ligament cells (지속적 및 간헐적 가압력이 치주인대 배양세포의 Alkaline Phosphatase 활성도에 미치는 영향)

  • Kwon, Suk-Yee;Bae, Seong-Min;Kyung, Hee-Moon;Sung, Jae-Hyun
    • The korean journal of orthodontics
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    • v.27 no.4 s.63
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    • pp.599-605
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    • 1997
  • The propose of this study was to evaluate the effect of cellular activity on PDL cells dependent on intermittent and continuous compressive force by determining the alkaline phosphatase activity. An intermittent and continuous compressive forces were applied on PDL cells at the confluent stage. The alkaline phosphatase activity was measured on control and experimental groups every 24, 48, 72hours. The experimental group were consist of continuous and intermittent compressive group which were compressed by $300g/cm^2$ of diaphram pump. The intermittent compressive group was connected by timer which was worked on 10 minutes and off 10minutes. The results were as follows ; 1. The alkaline phosphatase activity of intermittent compressive group was lower than control group at 24 hours(P<0.05). 2. The alkaline phosphatase activity between each groups showed no significant differences at 48hours. 3. The alkaline phosphatase activity of continuous compresssive group was significantly higher than control group at 72 hours(P<0.01).

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