• Animal cells and systems
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• v.14 no.2
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• pp.73-81
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• 2010

In this study, monoclonal antibodies against lysosomal acid phosphatase (LAP) of a salamander, Hynobius leechii, were used to determine the spatial and temporal expression of the LAP in the regenerating limbs. The Western blot and immunohistochemical analysis in the limb regeneration revealed that LAP was highly expressed at the dedifferentiation stage, especially in the wound epidermis and dedifferentiating limb tissues such as muscle and cartilage. With RA treatment, the LAP expression became upregulated in terms of both level and duration in the wound epidermis, blastemal cell and dedifferentiating limb tissues. In addition, in situ activity staining of LAP showed a similar result to that of immunohistochemistry. Thus, the activity profile of LAP activity coincides well with the expression profile of LAP during the dedifferentiation period. Furthermore, to examine the effects of lysosomal enzymes including LAP on salamander limb regeneration, lysosome extract was microinjected into limb regenerates. Interestingly, when the lysosome extract was microinjected into limb regenerates with a low dose of RA($50\;{\mu}g/g$ body wt.), skeletal pattern duplication occurred frequently in the proximodistal and transverse axes. Therefore, lysosomal enzymes might cause the regenerative environment and RA plays dual roles in the modification of positional value as well as evocation of extensive dedifferentiation for pattern duplication. In conclusion, these results support the hypothesis that dedifferentiation is a crucial event in the process of limb regeneration and RA-evoked pattern duplication, and lysosomal enzymes may play important role(s) in this process.

• 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 $Cl^-$.

• The Korean Journal of Zoology
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• v.37 no.1
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• pp.58-65
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• 1994

Phosphatase의 저해제로 알려진 okadaic acid(OA)가 한국산 개구리(북방산개구리 참개구리) 난자의 성숙에 미치는 효과를 조사하였다. 북방산개구리 난자에 약 50 nl의 okadaic acid(0 5-500 mM)를 미세주입한 후 18시간 배양한 결과 0.5 mM의 농도에서 부터 난자의 핵붕괴를 일으키기 시작하였다 동면초기에 처리한 progesterone에 반응하지 않는 난자들도 OA에 의하여 성숙을 일으켰으며. 이 성숙은 배양액에 첨가한 cycloheximide(10 mg/ml)에 영향을 받지 않았다 또한 OA의 처리를 받고 일정시간 배양한 난자의 세포질에는 미성숙 난자의 성숙을 유도하는 maturation promoting factor (MPF)의 활성이 생기었다 참개구리의 난자도 역시 OA에 의해 성숙이 유도되었으며, 주입 후 6시간에서부터 핵붕괴가 일어나기 시작하였다 참개구리에서도 OA의 처리가 MPF의 활성을 촉진하는 것을 세포질내에 Hl histone kinase의 활성도가 증가하는 것으로 확인할 수 있었다 참개구리에서도 OA에 의한 성숙은 cycloheximide나 CAMP에 의해 영향을 받지 않았다 이러한 결과들은 개구리 난자의 MPF 활성화와 성숙과정에 phosphatase가 관여함을 보여주는 것이다.

• Plant Biotechnology Reports
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• v.5 no.1
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• pp.45-51
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• 2011

A low acid phosphatase 3 (lap3) mutant was identified and characterized from an Arabidopsis activation-tagged (Weigel) population. The roots of the lap3 plants showed lower acid phosphatase (APase) activity compared to wild-type ones under low-Pi conditions ($10{\mu}M\;Pi$). Plasmid rescue experiments revealed that the activation-tagging vector was inserted into the intergenic region between At4g31540 and At4g31550 in the Arabidopsis genome. The genotypic segregation of the lap3 mutation was tightly linked with the phenotypic segregation of root APase activity in the prgeny of lap3. The transcript level of the At4g31520 (SDA1: SEVERE DEPOLYMERIZATION OF ACTIN 1), located 7.4 kb from the CaMV 35S enhancers in the lap3 mutant, was significantly reduced compared to that in the wild type. It was speculated that cellular actin polymerization may be involved in Pi acquisition in higher plants.

• Journal of Veterinary Clinics
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• v.7 no.1
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• pp.429-438
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• 1990

The cytochemical characteristics of the hematopoietic cells in blood and bone marrow from 3 clinically healthy dogs were examined using a battery of cytochemical stains. Alkaline phosphotase activity was demonstrated in eosinophilic series and occassionally in progranulocytes. A variety of cells exhibited acid phosphatase activity, but tartrate-resistant acid phosphatase activity was seen only in eosinophilic series. Peroxidase activity was observed in myeloblasts to segmented cells of granulocytic series and in monocytes. ${\alpha}$-naphthyl acetate esterase activity was found in monocytes and occassionally in lymphocytes. Naphthyl-AS-D-chloroacetate esterase marked neutrophilic series from myeloblasts to segmented cells. ${\beta}$-Glucuronidase activity was detected in a variety of cells except the cells of erythrocytic series. Periodic acid Schiff stain-positive granules were demonstrated in the neutrophilic and eosinophilic series from myelocytes to segmented cells and in monocytes and occassionally in lymphocytes. Sudan black B stain-positive granules marked granulocytic series from myeloblasts to segmented tells and monocytes.

• Journal of Radiation Protection and Research
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• v.18 no.2
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• pp.61-69
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• 1993

We examined various enzyme activity changes by intraperitoneal injection uranium in the carp liver. These enzyme activity changes can be used as biochemical indicators of internal exposure to uranium. The results were followings ; 1) Total protein concentration decreased by intraperitoneal injection in the carp liver. 2) Lysosomal acid pretense and ${\beta}-glucuronidase$ activities increased in the liver until sixth intraperitoneal injection of uranium, but Lysosomal acid phosphatase activities decreased in the liver until the sixth injection of uranium. 3) Alkaline phosphatase activities sharply increased and Glutamate oxaloacetate Transaminase activities steadily decreased in the liver until the sixth injection of uranium. 4) Creatine %kinase activities steadily decreased and malate dehydrogenase activities sharply decreased in the liver after the primary injection of uranium. Any malate dehydrogenase activities was not detected after sixth injection of uranium.

• Journal of The Korean Society of Grassland and Forage Science
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• v.17 no.3
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• pp.285-292
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• 1997

This study was conducted to obtain the transgenic Brnssica napus plants with tobacco Apase gene using the binary vector system of Agrobacteriurn fumefociens. The results obtained were summarized as follows: A repressible acid phosphatase gene of Saccharon~yces cerevisiae, pho105 was used for screening of tobacco Apase cDNA. In order to identify Apase gene in tobacco genome, Southern blot analysis was pcrformed and the Apase gcnc may be present as a single copy, or at most two or three copies, in tobacco genome. To isolate the tobacco Apase gene, tobacco cDNA library was constructed using purifed mRNA from -Pi treated tobacco root and the plaque forming unit of the library was 2.8 x $10^5$ pfu/m${\ell}$, therefore the library might cover all expressed mRNAs. Using pho5 as a probe. tobacco Apase cDNA was cloned, and restriction mapping and Southern blot analysis of cDNA insert were revealed that the 3.6 kb cDNA contained tobacco acid phosphatase cDNA. Plasmid pGA695 -tcAPl was constructed by subcloning tobacco Apase cDNA into the Hind site of pGA695 with 35s promoter which can be expressed constitutively in plants. The Brassica napus cotyledonary petioles were cocultivated with the ,4 grobacteriunz and transferred to the selection medium. The transformed and regenerated plants were transplanted to soil medium. Southern blot analysis was done on the transformed plants, and it was confirmed that a foregin gene was stably integrated into the genonies of B. nnpus plants.

• Mass Spectrometry Letters
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• v.9 no.3
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• pp.67-72
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• 2018

Alkaline phosphatase (AP) is a membrane-bound glycoprotein that is widely distributed in the plasma membrane of cells of various organs and also found in many organisms from bacteria to humans. The complete amino acid sequence and three-dimensional structure of human placental alkaline phosphatase have been reported. Based on the literature data, AP consists of two presumptive glycosylation sites, at Asn-144 and Asn-271. However, it only contains a single occupied N-linked glycosylation site and no occupied O-linked glycosylation sites. Hydrophilic interaction chromatography (HILIC) has been primarily employed for the characterization of the glycan structures derived from glycoproteins. N-glycan structures from human placental alkaline phosphatase (PLAP) were investigated using HILIC-Orbitrap MS, and subsequent data processing and glycan assignment software. 16 structures including 10 sialylated N-glycans were identified from PLAP.

• Archives of Pharmacal Research
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• v.22 no.5
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• pp.464-473
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• 1999

Regulation of phosphcholine-hydrolyzing phosphatase (phosphocholine-phosphatase) activity, purified from bovine brain, was examined under physiological conditions. Various endogenous phosphomonoesters, which were utilized as substrate, inhibited the phosphocoline-phosphatase activity competitively (Ki 5.5-$82.0 {\mu}M$); among phosphomonoesters tested, there was a similar order of capability between the binding affinity of substrate and the inhibitory potency. In addition, phosphate ions also inhibited the phosphatase activity competitively with a Ki value of approximately $16{\mu}M$. Although leucine or theophylline inhibited the phosphatase activity at pH 9.0, their inhibitory action decreased greatly at pH 7.4. The pH-Km and pH-Vm profiles indicate that ionizable amino acids are involved in substrate binding as well as catalysis, alluding that the phosphatase activity may be highly dependent on the intracellular pH. Amino acid modification study supports the existence of tyrosine, arginine or lysine residue in the active site, and the participation of tyrosine residue in the catalytic action may e suggested positively for the susceptibility to the action of tetranitromethane or HOl-generator. Separately, the oxidative inactivation of phosphocholine-phosphatase activity was investigated. Of oxidants tested, HOONO, HOCl, HOl and $ascorbate/Cu^{2+}$ system were effective to inactivate the phosphatase activity. Noteworthy, a remarkable inativation was accomplished by $30{\mu}M$ HOCl in combination with 1 mM Kl. Inaddition, $Cu^{2+}(3{\mu}M) $in combination with ascorbate at concentrations as low as 0.1-0.3 mM reduced the phosphatase activity to a great extent. From these results, it is proposed that the phosphocholine-phosphatase activity may be regulated endogenously and susceptible to the various oxidant system in vivo.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.33 no.2
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• pp.112-121
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• 1988

Soybean seed coat consisted of three layers, and the aleurone layer was attached to the innermost part of seed coat. It showed the different morphological characteristics with single cell layer compared with many cell layers in barley aleurone layer. The structural difference in aleurone cell among varieties was not detected in this experiment. The hole of middle part of hilum consisted of net formed material in order to pass water and gas. In the experiment, it was not studied whether the varieties with hard seed consist of the same structure or not, but the detailed study on this would be necessary. The activity of acid phosphatase showed a tendency to increase in process of imbibition in distilled water. There was no significant difference in the enzyme activity among the varieties tested, but the enzyme activity of Danyoupkong was slightly higher than that of Hwanggeumkong. In germinability, Danyoupkong is higher than Hwangkeumkong, so it might be attributed to the higher activity. There was no difference in acid phosphatase activity between released from the aleurone cell and accumulated in the aleurone cell from 6 to 12 hours incubation of the medium in the absence of GA$_3$, but the difference was detected after 12 hours incubation. And enzyme activity was the highest in the 18 hours incubation. The increase in the release of acid phosphatase from the aleurone cell would be considered as passive diffusive effect due to the increase in turgo pressure of aleurone cell. The acid phosphatase released from aleurone layer increased greatly after 24 hours incubation of the medium in the presence of GA$_3$ and the accumulation within the aleurone cell decreased linearly after 18 hours incubation. The result indicates that GA$_3$ enhance the rate of enzyme release from aleurone layer, suggests that the aleurone cell wall be digested by the introduction of GA and the digested wall act as the channels for enzyme release.