• Title/Summary/Keyword: Domain-specificity

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IMMUNOREGULATORY EFFECTS OF A MONOCLONAL ANTIBODY TO HUMAN 4-1 BB MOLECULE ON ALLOANTIGEN-MEDIATED IMMUNE RESPONSES.

  • Kim, Joong-Gon;Lee, Soo-Hyun;Lee, Jae-Woo;B.S. Kwon;Kang, Chang-Yuil
    • Proceedings of the Korean Society of Applied Pharmacology
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    • 1995.04a
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    • pp.82-82
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    • 1995
  • 4-lBB molecule is expressed on the surface of activated CD4$\^$+/ and CD8$\^$+/ T cells. We generated a panel of anti-4-1 B5 murine mAbs using a fusion protein consisting of the extracellular domain of human 4-1 BB fused to Glutathione S-transferase. The binding activity against cell surface 4-1 BB molecule was assessed by flow cytometry analysis. These studies showed that several anti-4-1 BB mAbs bound to 10-30% of CD4$\^$+/ and CD8$\^$+/T cells in PHA or Con A stimulated PBLs, although these mAbs interacted with only, l-2% of CD4$\^$+/ and CD8$\^$+/ T cells in normal PBLs, indicating the specificity of mAbs to the 4-l BB molecule on activated CD4$\^$+/ and CD8$\^$+/ T cells. Next, we examined the effect of an anti-4-l BB mAb (4B4-1-1) on allogeneic mixed lymphocyte reactions (MLRs). The data indicated that the antibody significantly inhibited the proliferative response at higher concentrations. When tested with several T cell mitogens, the antibody had no stimulatory or inhibitory effects on the mitogen-mediated T cell proliferation. These data suggest that 4-1 BB molecule may play a role in the regulation of antigen-mediated immune response.

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A Leveling and Similarity Measure using Extended AHP of Fuzzy Term in Information System (정보시스템에서 퍼지용어의 확장된 AHP를 사용한 레벨화와 유사성 측정)

  • Ryu, Kyung-Hyun;Chung, Hwan-Mook
    • Journal of the Korean Institute of Intelligent Systems
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    • v.19 no.2
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    • pp.212-217
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    • 2009
  • There are rule-based learning method and statistic based learning method and so on which represent learning method for hierarchy relation between domain term. In this paper, we propose to leveling and similarity measure using the extended AHP of fuzzy term in Information system. In the proposed method, we extract fuzzy term in document and categorize ontology structure about it and level priority of fuzzy term using the extended AHP for specificity of fuzzy term. the extended AHP integrates multiple decision-maker for weighted value and relative importance of fuzzy term. and compute semantic similarity of fuzzy term using min operation of fuzzy set, dice's coefficient and Min+dice's coefficient method. and determine final alternative fuzzy term. after that compare with three similarity measure. we can see the fact that the proposed method is more definite than classification performance of the conventional methods and will apply in Natural language processing field.

A novel F-box protein with leucine-rich repeats affects defecation frequency and daumone response in Caenorhabditis elegans

  • Kim, Sung-Moon;Jang, Sang-Ho;Son, Na-Rae;Han, Ching-Tack;Min, Kwan-Sik;Lee, Hak-Kyo;Hwang, Sue-Yun
    • Animal cells and systems
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    • v.16 no.4
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    • pp.280-288
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    • 2012
  • Targeted degradation of proteins through ubiquitin-mediated proteolysis is an important control mechanism in various cellular processes. The process of ubiquitin conjugation is achieved by three enzyme complexes, among which the ubiquitin ligase complex (E3) is in charge of substrate specificity. The SCF (SKP1-CUL1-F-box) family portrays the largest and the most characterized member of the E3 ligases. For each SCF complex, the ubiquitination target is recognized by the F-box protein subunit, which interacts with the substrate through a unique C-terminal domain. We have characterized a novel F-box protein CFL-1 that represents a single LRR-type F-box (FBXL) in the Caenorhabditis elegans genome. CFL-1 is highly homologous to FBXL20 and FBXL2 of mammals, which are known to regulate synaptic vesicle release and cell cycle, respectively. A green fluorescence protein (GFP)-reporter gene fused to the cfl-1 promoter showed restricted expression around the amphid and the anus. Modulation of CFL-1 activity by RNAi affected the time interval between defecations. RNAi-treated worms also exhibited reduced tendency to form dauer when exposed to daumone. The potential involvement of CFL-1 in the control of defecation and pheromone response adds to the ever expanding list of cellular processes controlled by ubiquitin-mediated proteolysis in C. elegans. We suggest that CFL-1, as a single LRR-type F-box protein in C. elegans, may portray a prototype gene exerting diverse functions that are allocated among multiple FBXLs in higher organisms.

Recombinant Protein Expression and Purification of the Human HMTase MMSET/NSD2

  • Morishita, Masayo;Mevius, Damiaan;Shen, Yunpeng;Di Luccio, Eric
    • Current Research on Agriculture and Life Sciences
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    • v.31 no.3
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    • pp.157-164
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    • 2013
  • Chromatin remodelers that include histone methyl transferases (HMTases) are becoming a focal point in cancer drug development. The NSD family of three HMTases, NSD1, NSD2/MMSET/WHSC1, and NSD3/WHSC1L are bona fide oncogenes found aberrantly expressed in several cancers, suggesting their potential role for novel therapeutic strategies. Several histone modifiers including HMTase have clear roles in human carcinogenesis but the extent of their functions and regulations are not well understood, especially in pathological conditions. The extents of the NSDs biological roles in normal and pathological conditions remain unclear. In particular, the substrate specificity of the NSDs remains unsettled and discrepant data has been reported. NSD2/MMSET is a focal point for therapeutic interventions against multiple myeloma and especially for t(4;14) myeloma, which is associated with a significantly worse prognosis than other biological subgroups. Multiple myeloma is the second most common hematological malignancy in the United States, after non-Hodgkin lymphoma. Herein, as a first step before entering a pipeline for protein x-ray crystallography, we cloned, recombinantly expressed and purified the catalytic SET domain of NSD2. Next, we demonstrated the catalytic activities, in vitro, of the recombinantly expressed NSD2-SET on H3K36 and H4K20, its biological targets at the chromatin.

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Novel Genetic Associations Between Lung Cancer and Indoor Radon Exposure

  • Choi, Jung Ran;Koh, Sang-Baek;Park, Seong Yong;Kim, Hye Run;Lee, Hyojin;Kang, Dae Ryong
    • Journal of Cancer Prevention
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    • v.22 no.4
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    • pp.234-240
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    • 2017
  • Background: Lung cancer is the leading cause of cancer-related death worldwide, for which smoking is considered as the primary risk factor. The present study was conducted to determine whether genetic alterations induced by radon exposure are associated with the susceptible risk of lung cancer in never smokers. Methods: To accurately identify mutations within individual tumors, next generation sequencing was conduct for 19 pairs of lung cancer tissue. The associations of germline and somatic variations with radon exposure were visualized using OncoPrint and heatmap graphs. Bioinformatic analysis was performed using various tools. Results: Alterations in several genes were implicated in lung cancer resulting from exposure to radon indoors, namely those in epidermal growth factor receptor (EGFR), tumor protein p53 (TP53), NK2 homeobox 1 (NKX2.1), phosphatase and tensin homolog (PTEN), chromodomain helicase DNA binding protein 7 (CHD7), discoidin domain receptor tyrosine kinase 2 (DDR2), lysine methyltransferase 2C (MLL3), chromodomain helicase DNA binding protein 5 (CHD5), FAT atypical cadherin 1 (FAT1), and dual specificity phosphatase 27 (putative) (DUSP27). Conclusions: While these genes might regulate the carcinogenic pathways of radioactivity, further analysis is needed to determine whether the genes are indeed completely responsible for causing lung cancer in never smokers exposed to residential radon.

Characterization of a GH8 β-1,4-Glucanase from Bacillus subtilis B111 and Its Saccharification Potential for Agricultural Straws

  • Huang, Zhen;Ni, Guorong;Zhao, Xiaoyan;Wang, Fei;Qu, Mingren
    • Journal of Microbiology and Biotechnology
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    • v.31 no.10
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    • pp.1446-1454
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    • 2021
  • Herein, we cloned and expressed an endo-β-1,4-glucanase gene (celA1805) from Bacillus subtilis B111 in Escherichia coli. The recombinant celA1805 contains a glycosyl hydrolase (GH) family 8 domain and shared 76.8% identity with endo-1,4-β-glucanase from Bacillus sp. KSM-330. Results showed that the optimal pH and temperature of celA1805 were 6.0 and 50℃, respectively, and it was stable at pH 3-9 and temperature ≤50℃. Metal ions slightly affected enzyme activity, but chemical agents generally inhibited enzyme activity. Moreover, celA1805 showed a wide substrate specificity to CMC, barley β-glucan, lichenin, chitosan, PASC and avicel. The Km and Vmax values of celA1805 were 1.78 mg/ml and 50.09 µmol/min/mg. When incubated with cellooligosaccharides ranging from cellotriose to cellopentose, celA1805 mainly hydrolyzed cellotetrose (G4) and cellopentose (G5) to cellose (G2) and cellotriose (G3), but hardly hydrolyzed cellotriose. The concentrations of reducing sugars saccharified by celA1805 from wheat straw, rape straw, rice straw, peanut straw, and corn straw were increased by 0.21, 0.51, 0.26, 0.36, and 0.66 mg/ml, respectively. The results obtained in this study suggest potential applications of celA1805 in biomass saccharification.

Roles of Carbohydrate-Binding Module (CBM) of an Endo-β-1,4-Glucanase (Cel5L) from Bacillus sp. KD1014 in Thermostability and Small-Substrate Hydrolyzing Activity

  • Lee, Jae Pil;Shin, Eun-Sun;Cho, Min Yeol;Lee, Kyung-Dong;Kim, Hoon
    • Journal of Microbiology and Biotechnology
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    • v.28 no.12
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    • pp.2036-2045
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    • 2018
  • An endo-${\beta}$-1,4-glucanase gene, cel5L, was cloned using the shot-gun method from Bacillus sp.. The gene, which contained a predicted signal peptide, encoded a protein of 496 amino acid residues, and the molecular mass of the mature Cel5L was estimated to be 51.8 kDa. Cel5L contained a catalytic domain of glycoside hydrolase (GH) family 5 and a carbohydrate-binding module family 3 (CBM_3). Chromatography using HiTrap Q and CHT-II resulted in the isolation of two truncated forms corresponding to 50 (Cel5L-p50) and 35 kDa (Cel5L-p35, CBM_3-deleted form). Both enzymes were optimally active at pH 4.5 and $55^{\circ}C$, but had different half-lives of 4.0 and 22.8 min, respectively, at $70^{\circ}C$. The relative activities of Cel5L-p50 and Cel5L-p35 for barley ${\beta}$-glucan were 377.0 and 246.7%, respectively, compared to those for carboxymethyl-cellulose. The affinity and hydrolysis rate of pNPC by Cel5L-p35 were 1.7 and 3.3 times higher, respectively, than those by Cel5L-p50. Additions of each to a commercial enzyme set increased saccharification of pretreated rice straw powder by 17.5 and 21.0%, respectively. These results suggest CBM_3 is significantly contributing to thermostability, and to affinity and substrate specificity for small substrates, and that these two enzymes could be used as additives to enhance enzymatic saccharification.

A Study on the Scope and Determinants of Electronic Collaboration based on IT in Interorganizational Relationships (기업간 거래에서 정보기술을 활용한 전자적 협력의 범위와 선행요인에 관한 연구)

  • Choi, Su-Jeong
    • Journal of Information Technology Applications and Management
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    • v.15 no.4
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    • pp.159-188
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    • 2008
  • This study suggests strategies which can enable to creation of new opportunities of competitive advantages while operating a long lasting and consistent business with major trading partners, based on interorganizational information systems (IOISs) specially established and installed for interorganizational transactions. Nowadays, IOISs based mechanism having been widely expanded as a conventional business infrastructure for the interorganizational transactions and/or exchanges, it is customary difficult to obtain any strongly sound advantage over the competitors who have adopted even the simplest deployment of the IOIS mechanisms. In this connection, this study intends to investigate the interorganizational collaborative activities conducted by under the auspicious of IOISs, focused on the prospect of the exploitation of IOISs rather than the implementation of the IOISs. In this study, we, firstly, suggest the concept of Electronic Collaboration which can be defined by the collaborative activities conducted by IOISs, compared to the ones conducted on off-line. In addition, we suggest the Electronic Collaboration as a multi-dimensional concept, constituted by three sub-constructs, the Electronic Information Sharing (EIS), the Electronic Joint Activity (EJA), and the construction of the Electronic Relational Knowledge Store (ERKS). Secondly, we empirically verify the effects of relational and environmental determinants on the Electronic Collaboration. In this study, the relational determinants relate to the variables created in interorganizational relationship like Trust, Influence, Relational Specific Asset-asset invested for the transaction-, and Continuity of the relationship. On the other hand, the environmental determinants relate to the variables surrounding the relationship which are difficult to control. We consider Product Complexity, Technological Uncertainty, and Market Variability as the domain of the environmental determinants. To test our hypotheses, we conducted both paper-based survey and online-based survey. After refining the data with missing responses, a total of 150 data was used for analysis. The results were as follows : Firstly, it is statistically significant that the Electronic Collaboration is composed of EIS, EJA, and ERKS. In particular, the results imply that the firms are able to accumulate relational knowledge base as well as to exchange information or knowledge, and to conduct joint activities through effort to further expand the Electronic Collaboration. Secondly, we have verified the individual effects of the relational and the environmental determinants on the Electronic Collaboration. Product Complexity has been revealed as the most influential variable affecting the Electronic Collaboration. Next, Interorganizational Trust and Technological Uncertainty, in that order, have been seen to have significant effects on the Electronic Collaboration. In other words, when products or services seem to be difficult to standardize, and the core technologies seem to rapidly change, the need for the Electronic Collaboration increase. In addition, the observation dictates that the interorganizational trust turns out to be a critical variable in building a relationship and in seeking further collaboration. The results, further, illustrate that the environmental determinants are relatively more effective than the relational determinants, which is not consistent with a few prior researches relational determinants emphasized. It is because this study doesn't consider the size of the firm. A few researchers have given an emphasis on the relational determinants like trust and influence, especially from the perspective of small firms in interorganizational relationship. However, in our study, where all the sizes of the firms are contained, electronic collaboration is considerably affected by the environmental determinants.

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Characterization of Pv92, a Novel Merozoite Surface Protein of Plasmodium vivax

  • Lee, Seong-Kyun;Wang, Bo;Han, Jin-Hee;Nyunt, Myat Htut;Muh, Fauzi;Chootong, Patchanee;Ha, Kwon-Soo;Park, Won Sun;Hong, Seok-Ho;Park, Jeong-Hyun;Han, Eun-Taek
    • Parasites, Hosts and Diseases
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    • v.54 no.4
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    • pp.385-391
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    • 2016
  • The discovery and understanding of antigenic proteins are essential for development of a vaccine against malaria. In Plasmodium falciparum, Pf92 have been characterized as a merozoite surface protein, and this protein is expressed at the late schizont stage, but no study of Pv92, the orthologue of Pf92 in P. vivax, has been reported. Thus, the protein structure of Pv92 was analyzed, and the gene sequence was aligned with that of other Plasmodium spp. using bioinformatics tools. The recombinant Pv92 protein was expressed and purified using bacterial expression system and used for immunization of mice to gain the polyclonal antibody and for evaluation of antigenicity by protein array. Also, the antibody against Pv92 was used for subcellular analysis by immunofluorescence assay. The Pv92 protein has a signal peptide and a sexual stage s48/45 domain, and the cysteine residues at the N-terminal of Pv92 were completely conserved. The N-terminal of Pv92 was successfully expressed as soluble form using a bacterial expression system. The antibody raised against Pv92 recognized the parasites and completely merged with PvMSP1-19, indicating that Pv92 was localized on the merozoite surface. Evaluation of the human humoral immune response to Pv92 indicated moderate antigenicity, with 65% sensitivity and 95% specificity by protein array. Taken together, the merozoite surface localization and antigenicity of Pv92 implicate that it might be involved in attachment and invasion of a merozoite to a new host cell or immune evasion during invasion process.

Characterization of Caveola-Vesicle Complexes (CVCs) Protein, PHIST/CVC-8195 in Plasmodium vivax

  • Wang, Bo;Lu, Feng;Han, Jin-Hee;Lee, Seong-Kyun;Cheng, Yang;Nyunt, Myat Htut;Ha, Kwon-Soo;Hong, Seok-Ho;Park, Won Sun;Han, Eun-Taek
    • Parasites, Hosts and Diseases
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    • v.54 no.6
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    • pp.725-732
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    • 2016
  • Plasmodium vivax produces numerous caveola-vesicle complex (CVC) structures beneath the membrane of infected erythrocytes. Recently, a member helical interspersed subtelomeric (PHIST) superfamily protein, $PcyPHIST/CVC-81_{95}$, was identified as CVCs-associated protein in Plasmodium cynomolgi and essential for survival of this parasite. Very little information has been documented to date about $PHIST/CVC-81_{95}$ protein in P. vivax. In this study, the recombinant $PvPHIST/CVC-81_{95}$ N and C termini were expressed, and immunoreactivity was assessed using confirmed vivax malaria patients sera by protein microarray. The subcellular localization of $PvPHIST/CVC-81_{95}$ N and C termini in blood stage parasites was also determined. The antigenicity of recombinant $PvPHIST/CVC-81_{95}$ N and C terminal proteins were analyzed by using serum samples from the Republic of Korea. The results showed that immunoreactivities to these proteins had 61% and 43% sensitivity and 96.9% and 93.8% specificity, respectively. The N terminal of $PvPHIST/CVC-81_{95}$ which contains transmembrane domain and export motif (PEXEL; RxLxE/Q/D) produced CVCs location throughout the erythrocytic-stage parasites. However, no fluorescence was detected with antibodies against C terminal fragment of $PvPHIST/CVC-81_{95}$. These results suggest that the $PvPHIST/CVC-81_{95}$ is localized on the CVCs and may be immunogenic in natural infection of P. vivax.