• 한국생물공학회:학술대회논문집
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• 2001.11a
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• pp.225-228
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• 2001

RhRnBp and Peysn5 are the proteins related to carbohydrate synthesis. RhRnBp originated form human was expressed as inclusion body in E. coli. Peysn5 originated form actinomadura was expressed as low level and inclusion body in E. coli. Ub, Trx, MalE and NusA is used as fusion partner to RhRnBp and Peysn5. The solubility of all fusion protein is NusA > MalE> Trx > Ub. Expression level of RhRnBp fusions in $37^{\circ}C$ is higher than that in $25^{\circ}C$. However in the case of Peysn5. Expression levels in $25^{\circ}C$ were higher. MalE fusion had highest activity in RhRnBp fusions. There were no activity in Peysn5.

• The Journal of Korean Medicine
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• v.22 no.1
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• pp.63-77
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• 2001

Objectives : In acute stage of CVA, many patients experience Bugibultong as a symptom. Daesiho-tang is one of the most useful prescriptions for constipated stroke patients. This study investigated the effects of Daesiho-tang extracts on hypoxic E18 cortical neuroblast. Methods : To evaluate the effect of Daesiho-tang extracts on enuronal death caused by hypocia the neuronal viability and protein expressions of NMDA receptor subunits, ${\alpha}CaMK$ II PDE2 PSD95 densin-180, $eEF-l{\alpha}$ and GFAP were investigated. Results : We guessed that Daesiho-tang extracts worked to protect against cell damage through effecting on the protein expression of PSD(post synatpic density), especially increase of ${\alpha}CaMK$ II, $eEF-l{\alpha}$ PDE2, PSD95 and densin-180 Conclusions : According to the above results, it is supposed that Daesiho-tang is clinically applicable to stroke patients in the acute stage.

• Proceedings of the Computational Structural Engineering Institute Conference
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• 2007.04a
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• pp.719-724
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• 2007

Many engineering problems often require the large amount of computing resources for iterative simulations of problems treating many parameters and input files. In order to overcome the situation, this paper proposes an e-Science based computational system. The system exploits the Grid computing technology to establish an integrated web service environment which supports distributed high throughput computational simulations and remote executions. The proposed system provides an easy-to-use parametric study service where a computational service includes real time monitoring. To verify usability of the proposed system, two kinds of applications were introduced. The first application is an Aerospace Integrated Research System (e-AIRS). The e-AIRS adapts the proposed computational system to solve CFD problems. The second one is design and optimization of protein 3-dimensional structures.

• Mass Spectrometry Letters
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• v.12 no.4
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• pp.163-171
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• 2021

Emiliania huxleyi is a marine phytoplankton that plays a critical role in global carbon and sulfur cycling. The genome of E. huxleyi has been sequenced, and an in-depth proteomic profile of this organism has been reported. This study analyzed the phosphoproteome of E. huxleyi and identified its changes under calcium-limited conditions. A TiO₂ microcolumn was used for phosphopeptide enrichment, followed by liquid chromatography-tandem mass spectrometry analysis. Overall, we identified 7,010 phosphorylated sites on 3,355 phosphopeptides associated with 2,929 phosphoproteins in E. huxleyi. Quantitative analysis revealed changes in the phosphoproteome in E. huxleyi when ambient conditions changed to calcium-limited conditions, notably the phosphorylation of some transporters was altered. This study provides an overview of protein phosphorylation in E. huxleyi and paves the way for further investigations of its biological functions.

• BMB Reports
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• v.51 no.11
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• pp.572-577
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• 2018

Recent studies showed that the PD-1/PD-L1 checkpoint blockade is a dramatic therapy for melanoma by enhancing antitumor immune activity. Currently, major strategies for the PD-1/PD-L1 blockade have mainly focused on the use of antibodies and compounds. Seeking an alternative approach, others employ endogenous proteins as blocking agents. The extracellular domain of PD-1 (ePD1) includes the binding site with PD-L1. Accordingly, we constructed a PD-1-based recombinantly tailored fusion protein (dFv-ePD1) that consists of bivalent variable fragments (dFv) of an MMP-2/9-targeted antibody and ePD1. The melanoma-binding intensity and antitumor activity were also investigated. We found the intense and selective binding capability of the protein dFv-ePD1 to human melanoma specimens was confirmed by a tissue microarray. In addition, dFv-ePD1 significantly suppressed the migration and invasion of mouse melanoma B16-F1 cells, and displayed cytotoxicity to cancer cells in vitro. Notably, dFv-ePD1 significantly inhibited the growth of mouse melanoma B16-F1 tumor cells in mice and in vivo fluorescence imaging showed that dFv-ePD was gradually accumulated into the B16-F1 tumor. Also the B16-F1 tumor fluorescence intensity at the tumor site was stronger than that of dFv. This study indicates that the recombinant protein dFv-ePD1 has an intensive melanoma-binding capability and exerts potent therapeutic efficacy against melanoma. The novel format of the PD-L1-blocked agent may play an active role in antitumor immunotherapy.

• Journal of Plant Biotechnology
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• v.40 no.3
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• pp.169-177
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• 2013

Recently, the number of people with diabetes is rapidly increasing, coupled with the fact that the insulin market is remarkably increasing. Therefore, molecular farming for plant-derived pharmaceutical protein production is reported as becoming more attractive than ever. In this study, we carried out experiments step by step for development of recombinant insulin constructs, which were transformed into E. coli system, in vitro transcription and translation system, and tobacco cells. At first, recombinant proinsulin protein was successfully produced in in vitro transcription and translation system with wheat germ extract. After which, recombinant construct of prominiinsulin encoded a fusion protein of 7.8 kDa with trypsin cleavage sites at N terminus and C terminus of minimized C-peptide was tried to in vitro expression using E.coli culture. After purification with His-tag column, the resulting recombinant prominiinsulin protein was processed with trypsin, and then checked insulin biosynthesis by SDS-PAGE and western blot analysis with anti-insulin monoclonal antibody. The immunoreactive product of trypsin-treated miniinsulin was identical to the predicted insulin hexamer. The construct of 35S promoter-driven preprominiinsulin recombinant gene with signal peptide region for ER-targeting and red fluorescence protein gene [N terminus ${\rightarrow}$ tobacco E2 signal peptide ${\rightarrow}$ B-peptide (1-29 AA) ${\rightarrow}$ AAK ${\rightarrow}$ A-peptide (1-21 AA) ${\rightarrow}$ RR ${\rightarrow}$ His6 ${\rightarrow}$ KDEL ${\rightarrow}$ C terminus] was transformed into BY-2 tobacco cells. A polypeptide corresponding to the 38-kDa molecular mass predicted for fusion protein was detected in total protein profiles from transgenic BY-2 cells by western analysis. Therefore, this recombinant preprominiinsulin construct can be used for generation of transgenic tobacco plants producing therapeutic recombinant insulin.

• Journal of Life Science
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• v.8 no.3
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• pp.263-271
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• 1998

One of the better-characterized transcription factor of E. coli is the cAMP receptor protein(CRP) and the CRP binds cAMP and DNA. The cAMP-CRP complex is involved in regulation of many genes at bacteria. The cAMP-CRP regulatory element represents, in some respects, a global regulatory network. The aim of this work was to study the structure and the mechanisms controlling the expression of CRP in Serratia marcescens. We have been get 5 different clones from Serratia which stimulated the cells to use maltose as a sole carbon source in E. coli TP2139. The crp gene clone, pCKB12, was confirmed by Southern hybridization with E. coli crp gene. The location of the crp gene was determined by construction subclones carrying various portions of pCKB12. To investigate the potential role of CRP in E. coli, lacZ fused plasmids were constructed and investigated the ${\beta}$-galactosidase activity of the fused plasmid. The Serratiamarcescens cAMP receptor protein can substitute the E. coli CRP in transcriptional activation at the lacZ gene. These results suggest that Serratia marcescens cAMP receptor protein complex functions to regulate several promoters in E. coli.

• Asian-Australasian Journal of Animal Sciences
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• v.13 no.5
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• pp.605-612
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• 2000

Four young swamp buffalo cows of similar age ranging in weight between 280 to 380 kg and trained to do physical work were used in a study to determine energy and protein requirements for draught using a $4{\times}4$ Latin square designed experiment. The experiment consisted of field trials employing 4 levels of work load, e.g. no work as control, and loads amounting 450 to 500 Newton (N) pulled continuously for 1, 2 and 3 h daily for 14 consecutive days. Cows were fed king grass (Penisetum purpuroides) ad libitum and were subjected to materials balance trials. Body composition was estimated in vivo by the body density method and daily energy expenditure (EE) was calculated from ME minus retained energy (RE). The results show that EE while not working ($EE_{resting}$) was $0.42kgW^{0.75}MJ/d$ and maintenance ME ($ME_m$) was $0.37kgW^{0.75}MJ/d$. ME requirement increased to 1.65 times maintenance for the work of 3 hours. The energy expended for doing exercise ($E_{exercise}$) was 9.56, 20.0 and 25.86 MJ/cow for treatments 1, 2 and 3 II, respectively. Fat retention was absent in all groups of working cows, but protein retention was only negative for cows undertaking 3 h work. The relationship between $E_{exercise}$ (MJ), work load (F, kN), work duration (t, h) and body mass (W, kg) was found to be: $E_{exercise}=(0.003F^{1.43}t^{0.93})/W^{0.09}MJ$. The maintenance requirement for digestible protein was $2.51kgW^{0.75}g/d$, whereas digestible protein for growth ($DP_{growth}$) and for work ($DP_{work}$) followed the equations: $DP_{growth}=[(258+1.25W^{0.75}){\Delta}Wkg/d]g$ and $DP_{work}=[12.59e^{0.95t}]g$, respectively The coefficients a, b and c for the calculation of $E_{exercise}$ components according to the Lawrence equation were found to be 2.56 J/kgW.m, 5.2 J/kg load carried.m and 0.29, respectively, thus efficiencies to convert ME into work were 0, 16.09, 27.3 and 32.44% for control, 1, 2 and 3 h/d work, respectively. ME and DP requirements for a 250 to 400 kg working buffalo cow allowing to growth up to 0.5 kg/d are presented.

• BMB Reports
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• v.44 no.10
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• pp.659-664
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• 2011

As part of the search for biologically active anti-osteoporotic agents that enhance differentiation and mineralization of osteoblastic MC3T3-E1 cells, we identified the ginsenoside Rh2(S), which is an active component in ginseng. Rh2(S) stimulates osteoblastic differentiation and mineralization, as manifested by the up-regulation of differentiation markers (alkaline phosphatase and osteogenic genes) and Alizarin Red staining, respectively. Rh2(S) activates p38 mitogen-activated protein kinase (MAPK) in time- and concentration-dependent manners, and Rh2(S)-induced differentiation and mineralization of osteoblastic cells were totally inhibited in the presence of the p38 MAPK inhibitor, SB203580. In addition, pretreatment with Go6976, a protein kinase D (PKD) inhibitor, significantly reversed the Rh2(S)-induced p38 MAPK activation, indicating that PKD might be an upstream kinase for p38 MAPK in MC3T3-E1 cells. Taken together, these results suggest that Rh2(S) induces the differentiation and mineralization of MC3T3-E1 cells through activation of PKD/p38 MAPK signaling pathways, and these findings provide a molecular basis for the osteogenic effect of Rh2(S).

• Korean Journal of Microbiology
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• v.21 no.3
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• pp.115-126
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• 1983

The results that the effect of 6 detergents on the structural changes and biochemical composition of bacterial membrane of Escherichia coli and Bacillus cereus are as follows ; 1. Population growth of the bacteria was increased in case of the treatment with palmitoyl carnitine and sodium deoxy cholate but was increased in case of the treatment with palmitoyl carnitine and sodium deoxy cholate but was decreased by sodium dodecyl sulfate and palmitoyl choline, in E.coli and was decreased by palmitoyl carnitine and palmitoyl choline at the low concentration, in B. cereus. 2. The electron micrograph showed that cell wall lysis or cell collapse were observed in the treatment of sodium dodecyl sulfate and palmitoyl choline, and also cell wall was condensed by triton X-100 and sodium deoxy cholate, in E.coli. And in B. cereus, endospore formation of the bacteria was stimulated by palmitoyl choline, and cell lysis or structural changes of the membrane were observed in the treatment of sodium dodecyl sulfate, sodium cholate, and triton X-100, respectively. 3. As to the effect of detergent on the biochemical composition of biomembrane, the content of carnitine, in E.coli, and B.cereus, the content of structural protein and phospholipid were decreased by treatment of sodium dodecyl sulfate and structural protein was denatured by palmitoyl choline. 4. The profile of membrane protein revealed that the bacterial membrane were composed of various proteins. By dint of this result, some of membrane proteins were solubilized or changed to small molecules by the treatment of sodium dodecyl sulfate and palmitoyl choline, in E.coli and membrane protein of the biomembrane by treatment of sodium dodecyl sulfate, sodium deoxy cholate, palmitoyl choline, and palmitoyl carnitine were confirmed to be different profile as compared with those of the control, in B. cereus. Therefore, it is suggested that sodium dfodecyl sulfate and palmitoyl choline soulbilized biomembranes or inhibited membrane transport and that palmitoyl carnitine and sodium deoxy cholate were used as an energy source or stimulating the membrane transport, in E.coli. And, it is suggested that all of detergents were inhibited biomembrane synthesis, expet saponin, in B.cereus.