• Title/Summary/Keyword: ShK

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Molecular Cloning and NMR Characterization of the Nonreceptor Tyrosine Kinase PTK6 SH3-SH2-Linker Domain

  • Lee, Young-Min;Ahn, Kyo-Eun;Ko, Sung-Geon;Lee, Weon-Tae
    • Bulletin of the Korean Chemical Society
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    • v.30 no.5
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    • pp.1043-1046
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    • 2009
  • Human protein tyrosine kinase-6 (PTK6) is a member of the non-receptor protein tyrosine kinase family and it is found in two-thirds of all breast tumors. Very recently, we proposed that the SH3 domain of PTK6 interacts with the linker region (Linker) between the SH2 and kinase domains, proving that the interaction between SH3 domain and Linker plays an important role in auto-inhibition mechanism. Residues from 1 to 191 corresponding region of SH3-SH2-Linker (SH32L) of PTK6 was cloned into the pET32a expression vector with Tobbaco etch virus (TEV) protease enzyme site by sequence homology and 3D structural model. The purified PTK6-SH32L was determined as a monomer conformation in solution. The amide proton resonances in the $^{15}N-^{1}H$ 2D-HSQC spectrum suggest that PTK6-SH32L possesses disordered structural region of the flexible/unstructured linker region. In addition, the backbone amide proton chemical shifts of the SH3 domain in the PTK6-SH32L differ from that of the independent domain, indicating that intra-molecular interaction between SH3 and Linker in the PTK6-SH32L is present.

Application of Chitosan Preparations for Eco-friendly Control of Potato Late Blight (감자 역병의 친환경 방제를 위한 키토산 제형의 살포)

  • Chang, Taehyun;Kim, Byung Sup
    • Research in Plant Disease
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    • v.18 no.4
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    • pp.338-348
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    • 2012
  • Potato late blight caused by Phytophthora infestans Cooke is one of the major diseases in the cultivation of potatoes in Korea. Effect of chitosan preparations (SH-1 and SH-2) was evaluated on the inhibition of mycelial growth of P. infestans, and protective activity using detached potato leaf assay both in vivo and in the field test. SH-1 and SH-2 were showed protective activity of young plant with control values more than 95% potato late blight by inoculation with pathogens under growth chamber conditions. Mycelial growth was inhibited the radial growth over 74% at a concentration of $300{\mu}g/ml$ of both SH-1 and SH-2. Spraying with SH-1 and SH-2 on the leaves for detached leaf assay reduced disease development. The content of total polyphenol in stem was significantly increased by SH-1 and SH-2 application in the field. In field experiments, foliar application with both SH-1 and SH-2 were significantly reduced the development of late blight on potato plants. Control of late blight disease was obtained with control values of 72% and 53% by application of 1% SH-1 and SH-2, respectively, with 4 times at 7 days interval, and reduced with similar disease control values by application with 3 times at 14 days interval compared with untreated control. SH-1 and SH-2 applications increased the fresh weight of potato, and higher grade potatoes were also increased. The results showed that SH-1 and SH-2 applications can be used as eco-friendly natural fungicide for organic farming for the increase of yields and control of late blight.

Thermostability of Chimeric Cytidine Deaminase Variants Produced by DNA Shuffling

  • Park, Yu-Mi;Phi, Quyet Tien;Song, Bang-Ho;Ghim, Sa-Youl
    • Journal of Microbiology and Biotechnology
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    • v.19 no.12
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    • pp.1536-1541
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    • 2009
  • The DNA shuffling technique has been used to generate libraries of evolved enzymes in thermostability. We have shuffled two thermostable cytidine deaminases (CDAs) from Bacillus caldolyticus DSM405 (T53) and B. stearothermophilus IFO12550 (T101). The shuffled CDA library (SH1067 and SH1077 from the first round and SH2426 and SH2429 from the second round) showed various patterns in thermostability. The CDAs of SH1067 and SH1077 were more thermostable than that of T53. SH2426 showed 150% increased halftime than that of T53 at $70^{\circ}C$. The CDA of SH2429 showed about 200% decreased thermostability than that of T53 at $70^{\circ}C$. A single amino acid residue replacement that presented between SH1077 and SH2429 contributed to dramatic changes in specific activity and thermostability. On SDS-PAGE, the purified CDA of SH1077 tetramerized, whereas that of SH2429 denatured and became almost monomeric at $80^{\circ}C$. A simulated three-dimensional structure for the mutant CDA was used to interpret the mutational effect.

SH2D4A regulates cell proliferation via the ERα/PLC-γ/PKC pathway

  • Li, Tingting;Li, Wei;Lu, Jingyu;Liu, Hong;Li, Yinghui;Zhao, Yanyan
    • BMB Reports
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    • v.42 no.8
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    • pp.516-522
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    • 2009
  • SH2D4A, comprising a single SH2 domain, is a novel protein of the SH2 signaling protein family. We have previously demonstrated SH2D4A is expressed ubiquitously in various tissues and is located in the cytoplasm. In this study we investigated the function of SH2D4A in human embryonic kidney (HEK) 293 cells using interaction analysis, cell proliferation assays, and kinase activity detection. SH2D4A was found to directly bind to estrogen receptor $\alpha$ (ER$\alpha$), and prevent the recruitment of phospholipase C-$\gamma$ (PLC-$\gamma$) to ER$\alpha$. Moreover, we observed its inhibitory effects on estrogen-induced cell proliferation, involving the protein kinase C (PKC) signaling pathway. Together, these findings suggested that SH2D4A inhibited cell proliferation by suppression of the ER$\alpha$/PLC-$\gamma$/PKC signaling pathway. SH2D4A may be useful for the development of a new anti-cancer drug acting as an ER signaling modulator.

MeSH Semi Indexing of the Korean Biomedical Literature, using NLM Medical Text Indexer (NLM Medical Text Indexer를 활용한 우리나라 의학문헌의 MeSH Semi Indexing 방안)

  • Jeong, Sona;Lee, Choon Shil
    • Proceedings of the Korean Society for Information Management Conference
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    • 2010.08a
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    • pp.21-28
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    • 2010
  • 본 연구에서는 PubMed에 등재되었으나 Medical Subject Headings(MeSH)가 부여되지 않은 국내 의학학술지의 문헌을 대상으로 미국국립의학도서관 (NLM: National Library of Medicine)의 Medical Text Indexer(MTI)를 활용하여 MeSH 용어를 추천받은 후, PubMed 레코드의 유사주제문헌 (Relation Citations, PRC)에 부여된 MeSH와의 일치여부를 분석하였다. 또한 논문의 저자가 부여한 키워드(저자키워드)와 PRC MeSH의 일치여부도 비교하였다. PRC MeSH와 MTI MeSH 추천어의 일치율은 주표목이 21.1%였고, 체크태그는 18.1%, 부표목은 16.5%로 나타났다. 우리나라 의학논문에 나타난 저자키워드의 중요한 특징은 MeSH 주표목 위주이고, 체크태그와 부표목은 거의 사용하지 않는 것이다. 따라서 저자키워드와 PRC MeSH 주표목과의 일치율은 23.4%에 이르지만, 체크태그와 부표목의 일치율은 각각 1%, 2.1%였다. 색인전문가가 통제어휘를 사용하여 색인하는 과정에서 PRC와 MTI의 MeSH 주표목과 저자키워드가 일치하는 용어를 주표목으로 부여하고, PRC와 MTI가 추천하는 체크태그와 부표목을 활용하는 등 국내 의학문헌의 MeSH 용어 부여 작업을 반자동화(semi-indexing)하면, 정확하고 신속한 MeSH 부여 작업이 가능할 것이다.

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Development of ELISA System for Screening of Specific Binding Inhibitors for Src Homology (SH)2 Domain and Phosphotyrosine Interactions

  • Lee, Sang-Seop;Lee, Kyung-Im;Yoo, Ji-Yun;Jeong, Moon-Jin;Park, Young-Mee;Kwon, Byoung-Mog;Bae, Yun-Soo;Han, Mi-Young
    • BMB Reports
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    • v.34 no.6
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    • pp.537-543
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    • 2001
  • In the present study, an in vitro ELISA system to assess the interaction between Src homology (SH)2 domains and phosphotyrosine that contain peptides was established using purified GST-conjugated SH2 proteins and synthetic biotinylated phosphotyrosine that contain oligopeptides. The SH2 domains bound the relevant phosphopeptides that were immobilized in the streptavidin-coated microtiter plate in a highly specific and dose-dependent manner. The epidermal growth factor receptor (EGFR)-, T antigen (T Ag)-, and platelet-derived growth factor receptor (PDGFR)-derived phosphopeptides interacted with the growth factor receptor binding protein (Grb)2/SH2, Lck/SH2, and phosphatidyl inositol 3-kinase (PI3K) p85/SH2, respectively. No cross-reactions were observed. Competitive inhibition experiments showed that a short phosphopeptide of only four amino acids was long enough to determine the binding specificity. Optimal concentrations of the GST-SH2 fusion protein and phosphopeptide in this new ELISA system for screening the binding blockers were chosen at 2nM and 500nM, respectively. When two candidate compounds were tested in our ELISA system, they specifically inhibited the Lck/SH2 and/or p85/SH2 binding to the relevant phosphopeptides. Our results indicate that this ELISA system could be used as an easy screening method for the discovery of specific binding blockers of protein-protein interactions via SH2 domains.

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Comparison of Key Words of the Journal of Korean Academy of Fundamentals of Nursing with MeSH (2003-2007) (기본간호학회지 게재 논문의 주요어와 MeSH 용어의 비교(2003-2007년))

  • Chaung, Seung-Kyo;Sohng, Kyeong-Yae;Kim, Kyung-Hee
    • Journal of Korean Academy of Fundamentals of Nursing
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    • v.15 no.4
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    • pp.558-565
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    • 2008
  • Purpose: The purpose of this study was to analyze how accurately authors of the Journal of Korean Academy of Fundamentals of Nursing used MeSH terms as key words. Method: A total of 724 key words used in the 225 papers of Journal of Korean Academy of Fundamentals of Nursing from 2003 to 2007 were compared with MeSH terms. Results: Fifty nine point eight percent of total key words were completely coincident with MeSH terms, 13.5% were entry terms, and 21.8% were not MeSH terms. The coincidence rates for 2003 and 2007 separately were 38.5% and 70.9%. Also, 25.3% of papers precisely used MeSH terms as key words and 8% did not use any MeSH terms. Conclusion: The results show that the coincidence rate of key words with MeSH terms was at a moderate level and gradually increased according to year. However, there is a need for us to understand MeSH more specifically and accurately.

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Increases in Doxorubicin Sensitivity and Radioiodide Uptake by Transfecting shMDR and Sodium/Iodide Symporter Gene in Cancer Cells Expressing Multidrug Resistance (다약제내성 암세포에서 shMDR과 Sodium/Iodide Symporter 유전자의 이입에 의한 Doxorubicin 감수성과 방사성옥소 섭취의 증가)

  • Ahn, Sohn-Joo;Lee, Yong-Jin;Lee, You-La;Choi, Chang-Ik;Lee, Sang-Woo;Yoo, Jeong-Soo;Ahn, Byeong-Cheol;Lee, In-Kyu;Lee, Jae-Tae
    • Nuclear Medicine and Molecular Imaging
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    • v.41 no.3
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    • pp.209-217
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    • 2007
  • Purpose: Multidrug resistance (MDR) of the cancer cells related to mdr1 gene expression can be effectively treated by selective short hairpin RNA for mdr1 gene (shMDR). Sodium/iodide symporter (NIS) gene is well known to have both reporter and therapeutic gene characteristics. We have co-transfected both shMDR and NIS gene into colon cancer cells (HCT15 cell) expressing MDR and Tc-99m sestamibi and I-125 uptake were measured. In addition, cytotoxic effects of doxorubicin and I-131 therapy were also assessed after transfection. Material and Methods: At first, shMDR was transfected with liposome reagent into human embryonic kidney cells (HEK293) and HCT cells. shMDR transfection was confirmed by RT-PCR and western blot analysis. Adenovirus expressing NIS (Ad-NIS) gene and shMDR (Ad-shMDR) were co-transfected with Ad-NIS into HCT15 cells. Forty-eight hours after infection, inhibition of P-gycoprotein (Pgp) function by shMDR was analyzed by a change of Tc-99m sestamibi uptake and doxorubicin cytotoxicity, and functional activity of induced NIS gene expression was assessed with I-125 uptake assay. Results: In HEK293 cells transfected with shMDR, mdr1 mRNA and Pgp protein expressions were down regulated. HCT15 cells infected with 20 MOI of Ad-NIS was higher NIS protein expression than control cells. After transfection of 300 MOI of Ad-shMDR either with or without 10 MOI of Ad-NIS, uptake of Tc-99m sestamibi increased up to 1.5-fold than control cells. HCT15 cells infected with 10 MOI of Ad-NIS showed approximately 25-fold higher I-125 uptake than control cells. Cotransfection of Ad-shMDR and Ad-NIS resulted in enhanced cytotoxic by doxorubicin in HCT15 cells. I-131 treatment on HCT15 cells infected with 20 MOI of Ad-NIS revealed increased cytotoxic effect. Conclusion: Suppression of mdr1 gene expression, retention of Tc-99m sestamibi, enhanced doxorubicin cytotoxicity and increases in I-125 uptake were achieved in MDR expressing cancer cell by co-transfection of shMDR and NIS gene. Dual therapy with doxorubicin and radioiodine after cotransfection shMDR and NIS gene can be used to overcome MDR.

The Equality of Key Words of the Journal of Korean Dental Society of Anesthesiology with Medical Subject Headings (MeSH) (2001-2014) (대한치과마취과학회지 게재 논문들의 핵심용어와 MeSH 용어의 일치도)

  • Shim, Youn-Soo;Kim, Ah-Hyeon;You, Yong-Ouk;Kim, Il-Ho;Yu, Song-Yi;Lee, Kwang-Seok;Jeong, Chae-Yul;Kim, Eun-Hee;Maeng, Sun-Woo;An, So-Youn
    • Journal of The Korean Dental Society of Anesthesiology
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    • v.14 no.3
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    • pp.143-149
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    • 2014
  • Background: The purpose of this study was to analyze the equality between key words used in the Journal of Korean Dental Society of Anesthesiology and Medical Subject Headings (MeSH). Methods: A total of 666 English key words in 187 papers (average 3.5 words in a paper) from 2001 to 2014 were eligible for this study. We classified them according to matched, and non-matched terms. After descriptive analysis, we assayed patterns of errors in using MeSH, and reviewed frequently used non-MeSH terms. Results: Fifty nine point six percent (59.6%) of total key words were completely coincident with MeSH terms, 40.39% were not MeSH terms. Conclusions: The results show that the coincidence rate of key words with MeSH terms was at a moderate level. However, there is a need for us to understand MeSH more specifically and accurately. Use of proper key words aligned with the international standards such as MeSH is important to be properly cited. The authors should pay attention and be educated on correct use of MeSH as key words.

Isolation and Identification of Wild Yeasts from Schizandra (Schizandra chinensis) for Wine Production and Its Characterization for Physicochemical and Sensory Evaluations (야생효모의 분리, 동정과 이를 이용한 오미자 발효주의 이화학 및 관능 특성의 비교)

  • Lee, Si-Hyung;Park, Hae-Kyung;Kim, Myung-Hee
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.39 no.12
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    • pp.1860-1866
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    • 2010
  • The purpose of this research was to characterize physicochemical properties and sensory evaluation of schizandra wines fermented by the yeasts, Sacchromyces cerevisiae SH8094 (S. cerevisiae SH8094) and Sacchromyces cerevisiae SH2855 (S. cerevisiae SH2855) isolated from schizandra fruits and stems and compare these results with the results from commercial activated yeast (Lalvin 1118) and a commercial schizandra wine. Three different schizandra wines fermented by S. cerevisiae SH8094, S. cerevisiae SH2855, and Lalvin 1118 showed similar results in pH and titratable acidity. On the other hand, the schizandra wines fermented by S. cerevisiae SH8094 and S. cerevisiae SH2855 showed high brix ($14^{\circ}$brix), low alcohol content (9%), and low yeasts count (4.1 log CFU/mL), compared with the schizandra wine fermented by Lalvin 1118. Both schizandra wines made with S. cerevisiae SH8094 and S. cerevisiae SH2855 showed higher scores in swallowing and overall acceptability than the schizandra wine made with Lalvin 1118. When compared with a commercial schizandra wine, the schizandra wine fermented with S. cerevisiae SH8094 showed better qualities in aroma ($6.65{\pm}1.47$), color ($7.53{\pm}1.14$), and overall acceptability ($6.76{\pm}1.03$). In conclusion, S. cerevisiae SH8094 which was isolated from schizandra fruits and stems has a high potential in schizandra wine fermentation.