• Title/Summary/Keyword: PEP-1

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An Efficient System for the Expression and Purification of Yeast Geranylgeranyl Protein Transferase Type I

  • Kim, Hyun-Kyung;Kim, Young-Ah;Yang, Chul-Hak
    • BMB Reports
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    • v.31 no.1
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    • pp.77-82
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    • 1998
  • To purify the geranylgeranyl protein transferase type I (GGPT-I) efficiently, a gene expression system using the pGEX-4T-1 vector was constructed. The cal1 gene, encoding the ${\beta}$ subunit of GGPT-I, was subcloned into the pGEX-4T-1 vector and co-transformed into E. coli cells harboring the ram2 gene, the ${\alpha}$ subunit gene of GGPT-I. GGPT-I was highly expressed as a fusion protein with glutathione S-transferase (GST) in E. coli, purified to homogeneity by glutathione-agarose affinity chromatography, and the GST moiety was excised by thrombin treatment. The purified yeast GGPT-I showed a dose-dependent increase in the transferase activity, and its apparent $K_m$ value for an undecapeptide fused with GST (GST-PEP) was $0.66\;{\mu}M$ and the apparent value for geranylgeranyl pyrophosphate (GGPP) was $0.071\;{\mu}M$.

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Nucleotide Activation of Catabolic Threonine Dehydratase from Serratia marcescens (뉴클레오타이드에 의한 Serratia marcescens Catabolic Threonine Dehydratase의 활성화)

  • Choi, Byung-Bum
    • The Korean Journal of Food And Nutrition
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    • v.23 no.2
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    • pp.171-177
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    • 2010
  • The catabolic threonine dehydratase from Serratia marcescens ATCC 25419 was purified to homogeniety using Sephadex G-200 gel filtration and AMP-Sepharose 4B affinity chromatography. The molecular weight of the native enzyme was 120,000 by native pore gradient PAGE. The enzyme was composed of four identical subunits with subunit molecular weights of 30,000 by SDS-PAGE. The Km values of the enzyme for L-threonine with and without AMP were 7.3 and 92 mM, respectively. There were 2 moles of pyridoxal phosphate and 16 moles of free -SH groups per 1 mole of enzyme. The enzyme was inhibited by $\alpha$-ketobutyrate, pyruvate, glyoxylate, and phosphoenol pyruvate(PEP) in the presence of AMP, yet stimulated by cAMP and ADP. For enzyme properties in comparison with S. marcescens, E. coli, and S. typhimurium enzyme, such as the PLP content, number of free sulfhydryl groups, and existence of ADP binding site, the S. marcescens enzyme was more similar to the S. typhimurium enzyme than the E. coli enzyme. Of the three enteric bacteria, the E. coli and S. typhimurium enzyme was increased the activity by ADP and cAMP, respectively, but only the S. marcescens enzyme was increased the activity by both ADP and cAMP. Therefore, the subtle differences in the properties between enzymes from the three enteric bacteria may represent minor structural differences among these enzymes and warrants further study.

The Effects on Respiratory Strength Training on Respiratory Function and Trunk Control in Patient with Stroke (호흡 강화 훈련이 뇌졸중 환자의 호흡기능 및 체간조절 능력에 미치는 효과)

  • Kim, Min-Hwan;Lee, Wan-Hee;Yun, Mi-Jung
    • The Journal of Korean Physical Therapy
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    • v.24 no.5
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    • pp.340-347
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    • 2012
  • Purpose: This study was conducted in order to investigate the effect on respiratory function, trunk control, and functional activities of daily living (ADL) through respiratory muscle strength training in patients with chronic stroke. Methods: Eighteen subjects who were six months post stroke participated in this study. The subjects were randomly allocated to two groups: experimental (n=10) and control (n=8). Both groups received physical therapy for five sessions, 30 minutes per week, during a period of six weeks. Subjects in the experimental group participated in an additional respiratory muscle strength training program, in which the threshold PEP device was used for 30 minutes per day, three days per week, during a period of six weeks. Results from pretest-posttest control were evaluated by pulmonary function forced vital capacity (FVC), forced expiratory volume at one second (FEV1), FEV1/FVC, peak expiratory flow (PEF), capacity of cough, trunk control, and functional ADL. Results: Significant improvement in the FEV1, PEF of pulmonary function (p<0.05), cough function (p<0.05), and trunk control (p<0.05) was observed among the groups. Conclusion: Respiratory muscle strength training improves pulmonary function, capacity of cough, and trunk control. These results suggest that respiratory muscle strength training is feasible and suitable for individuals with chronic stroke.

Inhibition of Prolyl 4-Hydroxylase by Oxaproline Tetrapeptides In Vitro and Mass Analysis for the Enzymatic Reaction Products

  • Moon Hong-sik;Begley Tedhg P.
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.5 no.1
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    • pp.61-64
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    • 2000
  • A series of 5-oxaproline peptide derivatives was synthesized and evaluated for its ability to inhibit the prolyl 4-hydroxylase in vitro. Structure-activity studies show that the 5-oxaproline sequences, prepared by the 1,3-dipolar cycloaddition of the C-methoxycarbonyl-N-mannosyl nitrone in the presence of the ethylene, are more active than the corresponding proline derivatives. Prolyl 4-hydroxylase belongs to a family of $Fe^{2+}-dependent$ dioxygenase, which catalyzes the formation of 4-hydroxyproline in collagens by the hydroxylation of proline residues in -Gly-Xaa-Pro-Gly- of procollagen chains. In this paper we discover the more selective N-Cbz-Gly-Phe-Pro-Gly-OEt $(K_m\;=\;520\;{\mu}M)$ sequences which are showed stronger binding than others in vitro. Therefore, we set out to investigate constrained tetrapeptide that was designed to mimic the proline structure of pep tides for the development of prolyl 4-hydroxylase inhibitor. From this result, we found that the most potent inhibitor is N-Dansyl-Gly-Phe-5-oxaPro-Gly-OEt $(K_i\;=\;1.6\;{\mu}M)$. This has prompted attempts to develop drugs which inhibit collagen synthesis. Prolyl 4-hydroxylase would seem a particularly suitable target for antifibrotic therapy.

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Subjective Symptoms of Cumulative Trauma Disorder in Furniture Manufacturing Workers (가구제조업 근로자의 누적외상성장해에 대한 자각증상)

  • Lee, Kyoung Ok
    • Korean Journal of Occupational Health Nursing
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    • v.6 no.1
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    • pp.44-52
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    • 1997
  • This study was carried out from January to March in 1996 to investigate subjective symptoms of cumulative trauma disorder (CTD). This survey was done by questionnaire of the Personal Ergonomics Profiles (PEP), which was developed by Johnson & Johnson. Number of respondents was 365 employees who were working in three furniture manufactories in Kwangjoo at Province Kyungki. The results were as follows : 1. There was no difference between clerical and labor workers in pain frequencies classified by general characteristics. However, labor workers showed statistically significant differences between working process in pain frequencies. Employees undertaking lifting work showed highest scores in frequencies of pain complaints. Comparing the number of pain occurrences, the married and the female was resulted in high score than the single and the male. There was no differences between ages and working experiences in group. 2. According to the mean frequency of body pain, the back was the most part to be complained in 1. 16 and followed by shoulder, neck, knee, wrist/hand, and forearm in order. Although, the study results showed that the workers working in furniture manufacture were not shown that they have had severe CTD, the female, the married and the the lifting workers were indicated as risk groups to CTD. Therefore, the development of backache prevention programe is necessary to manage pain control and to reduce health risks of CTD employees in their work environment.

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2007-2011 Characteristics of Plant Virus Infections on Crop Samples Submitted from Agricultural Places (2007-2011 우리나라 농업현장 임상진단 요청 작물의 바이러스 감염 특성)

  • Kim, Jeong-Soo;Lee, Su-Heon;Choi, Hong-Soo;Kim, Mi-Kyeong;Kwak, Hae-Ryun;Kim, Jeong-Sun;Nam, Moon;Cho, Jeom-Deog;Cho, In-Sook;Choi, Gug-Seoun
    • Research in Plant Disease
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    • v.18 no.4
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    • pp.277-289
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    • 2012
  • The total number of requests and associated specimens for the diagnosis of virus infection were 573 and 2,992, respectively, on crops from agricultural places of farmers, Agricultural extension services and so forth for 5 years from 2007. The total number of virus tests was 13,325. The number of species of viruses infected on the submitted crops was 21 in 2007, 15 in 2008, 23 in 2009, 21 in 2010 and 17 in 2011. The newly recorded viruses were Tobacco leaf curl virus (TbLCV) in 2007, Tomato yellow leaf curl virus (TYLCV) in 2008, Impatience necrotic spot virus (INSV) and Radish mosaic virus (RaMV) in 2009, and Beet western yellows virus (BWYV) in 2010. Forty virus species including Alfalfa mosaic virus were detected over 5 years. The ten most frequently detected virus species were Cucumber mosaic virus (CMV), Tomato spotted wilt virus (TSWV), Tomato leaf curl virus (TYLCV), Cucumber green mottle mosaic virus (CGMMV), Broad bean wilt virus 2 (BBWV2), Zucchini yellow mosaic virus (ZYMV), Melon necrotic spot virus (MNSV), Pepper mild mottle virus (PMMoV), Watermelon mosaic virus (WMV) and Pepper mottle virus (PepMoV). The types of crops submitted from agricultural places were 51 in total and the ten most frequently submitted crops were red pepper, tomato, paprika, watermelon, melon, rice, cucumber, corn, radish and gourd. The total request rate for the top 10 crops and top 20 crops was 81.6% and 94.2%, respectively. Eight pepper infecting virus species included CMV, and the average infection rate was 24.6% for CMV, 18.9% for PMMoV and 14.7% for TSWV. Seven kinds of double infection were detected in pepper including BBWV2+CMV at 14.7% on average, and four types of triple infection including BBWV2+CMV+PepMoV at 0.9% on average. Six virus species detected on tomato including TYLCV, and the average infection rate was 50.6% for TYLCV, 14.5% for TSWV and 10.9% for Tobacco leaf curl virus (TbLCV). The mixed infection of CMV+TSWV on tomato was 3.9% on average and of Tomato mosaic virus (ToMV)+TYLCV was 0.4% on average. Five viruses detected on watermelon included MNSV and the average infection rate was 37.0% for MNSV, 20.4% for CGMMV, 18.1% for ZYMV and 17.8% for WMV. The mixed infection rate on watermelon was CMV+MNSV and WMV+ZYMV having an average infection rate of 0.7% and 5.0%, respectively. The average infection rates on melon were 77.6% for MNSV, 5.6% for CMV and 3.3% for WMV. Mixed infections of CMV+MNSV occurred on melon with an average infection rate of 13.5%.

Survey and identification of virus diseases on paprika in Jeonnam province

  • Ko, Sug-Ju;Lee, Yong-Hwan;Cha, Kwang-Hong;An, U-Yup;Park, Hong-Soo
    • Proceedings of the Korean Society of Plant Pathology Conference
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    • 2003.10a
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    • pp.149.2-150
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    • 2003
  • Occurrences of virus diseases on paprika ( Capsicum annuum var. grossum) were surveyed in Joennam province from 1999 to 2003 and the collected samples showing virus-like symptoms were tested using ELISA. Virus diseases appeared 4.5%, 17.5%, and 4.9% in 2000, 2002, and 2003, respectively. As the results of investigation of the seasonal incidence with the growing stages of plant, virus was not occurred at seedling stage and was slightly from the planting time to the first harvesting time, but was dramatically increased at the second harvesting time. Virus diseases were more severe on the vinyl house than on the green house. Pepper mild mottle virus (PMMoV) was severely occurred in 2000 but not after that year. Comparing the virus species, Pepper mottle virus (PepMoV) was 35.9%, Broad bean wilt virus (BBWV) was 14.1%, and Cucumber mosaic virus (CMV) was 10.9% in 2002, and 76.0%, 11.1%, and 2.4% in 2003, respectively.

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Synthesis of Aminosilane treated Waterborne Poly Urethane/Epoxy Hybrid Resin used for Loudspeaker Damper (스피커 댐퍼용 아미노실란 처리 수분산 폴리우레탄/에폭시 하이브리드 수지의 합성)

  • Choi, Hyun-Seuk;Choi, Dong-Ho;Park, Ju-Yong;Huh, Man-Woo
    • Textile Coloration and Finishing
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    • v.29 no.1
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    • pp.25-36
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    • 2017
  • This study worked on the synthesis of waterborne polyurethane/epoxy hybrid resins containing aminosilane compound to replace conventional phenol resins used for a coating material for loudspeaker dampers, which are not harmful to human being. Waterborne polyurethane resins were synthesized from two diisocyanate of 4,4'-diphenylethane diisocyanate and toluene diisocyanate, two polyols of polyester polyols(PEP), polycarbonatediol(PCD), including and anionic center of dimethylol butanoic acid, a chain extenders of ethylenediamine(EDA), and a neutral agent of triethylamine. Synthesized polyurethane resins and commercially available bisphenol A type waterborne epoxy resin were blended in weight ratios of 80:20 to prepare polyurethane/epoxy hybrid resins. The synthesized waterborne polyurethane/epoxy hybrid resins were reacted with aminosilane compound to improve mechanical properties. Aminosilane-treated polyurethane /epoxy hybrid resins showed better mechanical properties.

Inhibitory Mechanism of Novel Inhibitors of UDP-N-Acetylglucosamine Enolpyruvyl Transferase from Haemophilus influenzae

  • Jin, Bong-Suk;Han, Seong-Gu;Lee, Won-Kyu;Ryoo, Sung-Weon;Lee, Sang-Jae;Suh, Se-Won;Yu, Yeon-Gyu
    • Journal of Microbiology and Biotechnology
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    • v.19 no.12
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    • pp.1582-1589
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    • 2009
  • Bacterial UDP-N-acetylglucosamine enolpyruvyl transferase (MurA) catalyzes the transfer of enolpyruvate from phosphoenolpyruvate (PEP) to uridine diphospho-N-acetylglucosamine (UNAG), which is the first step of bacterial cell wall synthesis. We identified thimerosal, thiram, and ebselen as effective inhibitors of Haemophilus influenzae MurA by screening a chemical library that consisted of a wide range of bioactive compounds. When MurA was preincubated with these inhibitors, their 50% inhibitory concentrations ($IC_{50}s$) were found to range from 0.1 to $0.7\;{\mu}M$. In particular, thimerosal suppressed the growth of several different Gram-negative bacteria such as Escherichia coli, Pseudomonas aeruginosa, and Salmonella typhimurium at a concentration range of $1-2\;{\mu}g/ml$. These inhibitors covalently modified the cysteine residue near the active site of MurA. This modification changed the open conformation of MurA to a more closed configuration, which may have prevented the necessary conformational change from occurring during the enzyme reaction.

Effects of Risering Design and Chemical Composition on Formation of Shrinkage Cavity in Gray Cast Iron (회주철의 수축결함생성에 미치는 주조방안 및 화학조성의 영향)

  • Yu, Sung-Kon
    • Korean Journal of Materials Research
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    • v.13 no.5
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    • pp.297-302
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    • 2003
  • The effects of risering design and alloying element on the formation of defects such as external depression, primary and secondary shrinkage cavity in gray cast iron were investigated. Two types of risering design for the cylindrically step-wise specimen, No. 1(progressive solidification) and No. 2(directional solidification) risering designs, were prepared and five different alloy compositions were casted. In the No. 1 risering design, external depression or primary shrinkage cavity due to liquid contraction was observed in all the specimens from ISO 150 to ISO 350. The primary shrinkage cavity was located right under the top surface or connected to the top surface, and was characterized by smooth surface. Its size increased with an increase in ISO number. However, neither secondary shrinkage cavity nor swollen surface was observed in all the castings. In the No.2 risering design, neither primary shrinkage cavity nor secondary shrinkage cavity was observed in all the specimens due to proper risering design. A swollen surface was also not observed in all the castings with the application of pep-set mold.