• Title/Summary/Keyword: Purification function

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Structural Characterization of Growth-Related Translationally Controlled Tumor Protein P23

  • Lee, Bong-Jin;Hong, Yoon-Hun;Park, Sang-Ho;Lee, Kyunglim
    • Journal of the Korean Magnetic Resonance Society
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    • v.5 no.1
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    • pp.46-55
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    • 2001
  • P23, a translationally controlled turner protein is involved in the interleukin-4 secretion from human basophils and is also known to be an IgE-dependent histamine-releasing factor. However, the precise physiological function and structure of P23 have not been elucidated. In the current study, we constructed the optimal expression and purification protocol of P23 and investigated the secondary structure and structural stability in various conditions. Circular dichroism (CD) investigation showed that the secondary structure of P23 adopts mainly a P-sheet conformation. CD spectroscopy and differential scanning calorimetry revealed that P23 is fairly stable in the pH range of neutral and mild-basic conditions and in the temperature range of 10 - 50$\^{C}$. Since the thermal stability and the P-sheet content of P23 were decreased by the addition of Ca$\^$2+/ ion, it could be suggested that Ca$\^$2+/ion induces structural change by partially destabilizing the structure of P23. In addition various H experiments were monitored to solve the aggregation of P23. Den results will provide the preliminary structural information about P23.

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Optimization of the experimental conditions for structural studies of the second transmembrane domain from human wild-type & mutant melanocortin-4 receptor

  • Gang, Ga-Ae;Choi, Sung-Sub;Park, Tae-Joon;Kim, Yong-Ae
    • Journal of the Korean Magnetic Resonance Society
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    • v.14 no.2
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    • pp.88-104
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    • 2010
  • Human melanocortin-4 receptor (hMC4R) has a critical role in part of energy homeostasis, and their heterozygous mutations related in genetic cause of severe human obesity. In order to study the structure and function of these membrane proteins, it is important to prepare the samples. However, the preparation of transmembrane peptide is seriously difficult and time-consuming. Overexpression and purification of membrane proteins was reported to be difficult due to their innate insoluble and toxic properties. Among the many difficulties, the most important is the difficulty in obtaining sufficient quantities of purified protein. Recently, we succeed to produce large amounts of the second transmembrane domain from the wild-type hMC4R (wt-TM2) and D90N mutant hMC4R (m-TM2) and proposed the structural difference of them in membrane-like environments. In this paper, we demonstrate the optimization procedures to express and purify wt-TM2 or m-TM2 peptides, and solution NMR studies in different detergents to get high-resolution spectra were also described.

Analysis of Purification Capacity of Paddy Fields Using Nutrient Balance (논에서 영양물지 수지를 이용한 논 정화능력 분석)

  • Jung, Jae Woon;Yoon, Kwang Sik;Choi, Woo Jung;Choi, Woo Young;Lee, Soo Hyung;Chang, Nam Ik;Hong, Soon Kang;Joo, Seuk Hun
    • Journal of Wetlands Research
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    • v.10 no.3
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    • pp.1-7
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    • 2008
  • In this study, the nutrient removal capacity of paddy field was investigated. Paddy field was considered as a constructed wetland. The present study was conducted during a cropping period in a rice cultivation area located in Hampyung-gun, Jeollanamdo, Korea. Water balance and mass balance of nutrients were estimated after intensive field monitoring. The surface runoff losses of T-N and T-P were 20.82 kg/ha and 1.55 kg/ha, respectively. The losses were 13.2% and 6.4% of T-N and T-P input into paddy fields, respectively. The results showed that the paddy fields have nutrient removal function.

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Separating and purifying polysaccharide, polypeptide from sea cucumber and their functions (해삼으로부터 다당류와 폴리펩타이드의 분리, 정제 및 기능성)

  • Jin, Hai-zhu;Fu, Xue-jun;Shen, Jing-yu;Sun, Bo;Wang, Hong-tao
    • Proceedings of the Korean Society of Postharvest Science and Technology of Agricultural Products Conference
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    • 2005.09a
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    • pp.112-123
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    • 2005
  • For thestudying and utilizing the thorn sea cucumber synthetically, the method preparing polysaccharide and polypeptide from sea cucumber was studied. The comparatively rational craft for preparing crude polysaccharide is fresh raw materials deal with oar form, $60\%$ of the ethanol precipitate after hydrolyzed by pretense and vibrated with ultrasonic wave auxiliary. The purification of polysaccharide and removal of protein fromcrude polysaccharide were made through precipitation method using the acetate. The polypeptide is obtained by concentrating in vacuum, freeze-drying after mixing precipitations of two times. Polysaccharide acute poisoning experiment indicate there is no bad reaction when LD50>5000g/kg, there strain rate of liver tumor H11 approach present generally acknowledged tumor treat medicine cyclophosphamide, and there is no side effect. The fatigue resistance function of polypeptide experiment also indicates that the fatigue resistance ability of mouse which fed on added sea cucumber polypeptide has a great improvement.

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Structural Effects of the GXXXG Motif on the Oligomer Formation of Transmembrane Domain of Syndecan-4

  • Song, Jooyoung;Kim, Ji-Sun;Choi, Sung-Sub;Kim, Yongae
    • Bulletin of the Korean Chemical Society
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    • v.34 no.12
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    • pp.3577-3585
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    • 2013
  • Syndecan-4 (heparan sulfate proteoglycan), biologically important in cell-to-cell interactions and tumor suppression, was studied through mutation of the GXXXG motif of its transmembrane domain (Syd4-TM), a motif which governs dimerization. The expression and purification of the mutant (mSyd4-TM) were optimized here to assess the function of the GXXXG motif in the dimerization of Syd4-TM. mSyd4-TM was obtained in M9 minimal media and its oligomerization was identified by SDS PAGE, Circular Dichroism (CD) spectroscopy, mass spectrometry and NMR spectroscopy. The mutant, unlike Syd4-TM, did not form dimers and was observed as monomers. The GXXXG motif of Syd-4TM was shown to be an important structural determinant of its dimerization.

A Numerical Analysis of the Abatement of VOC with Different Photocatalytic Honeycomb Filters (광촉매 필터형상에 따른 휘발성 유기화합물의 제거에 관한 수치해석적 연구)

  • 류무성;김창녕
    • Korean Journal of Air-Conditioning and Refrigeration Engineering
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    • v.16 no.1
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    • pp.1-7
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    • 2004
  • This study has been numerically conducted to investigate the removal efficiency of Volatile Organic Compound (VOC) for different photocatalytic honeycomb filters. Recently, the photocatalysis is being applied to air-cleaner, air-conditioner and vacuum-cleaner with the capability of air-purification, sterilization and antibiosis. However, photocatalysis is less efficient than other methods for removing VOC except in the case of low concentration. So far most of studies have focused on an improvement of the photocatalytic materials, but this study have placed emphasis on the improvements of shape of photocatalytic honeycomb filter. UV irradiation, concentration profile and pressure drop have been investigated for different cross sections of filters and for different filter lengths. Light intensity is dropped sharply with increasing distance from the UV-lamp, and becomes very low in the middle of the filters. Since photocatalytic reaction rate is a function of light intensity, VOC concentration gradient might be small in the middle of long filters. Thus, most of reaction have risen within only three times of dimensionless axial distance. These results can be used effectively for the design of advanced photocatalytic honeycomb filters.

Theoretical Consideration of the Plan for Environmental-Friendly Applications of Flood Plain around Dam (댐주변 범람지의 환경친화적 활용방안에 관한 이론고찰)

  • Shin, Byung-Chuel;Lee, Eun-Yeob
    • Journal of Environmental Science International
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    • v.12 no.2
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    • pp.185-193
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    • 2003
  • The purpose of this study was attempted to establish concepts of environmental-friendly applications of flood plain and to suggest the application plans. The results of this study can be summarized as follows; 1. Roles of flood plain as biotop (restoration, preservation, and creation of stream corridor ecosystem) should be considered. 2. Application methods considering environmental and scenic values should be reviewed. 3. Application methods reflecting values as regional ecological resources should be planned. 4. Preservation and application should be considered together, and obtaining a means of living for regional residents and creation of economic profits should be considered together. 5. Land application and approach method by usages (integrated management model) should be applied to utilize and manage flood plain efficiently. 6. Flood plain application programs should be designed reflecting opinions of regional residents. 7. With respect to space planning of flood plain, introduction of facilities focused on ecosystem preservation/ecosystem restoration/experiences/observation/learning/culture/ recreation/water purification could be reviewed positively.

High Level Expression of a Protein Precursor for Functional Studies

  • Gathmann, Sven;Rupprecht, Eva;Schneider, Dirk
    • BMB Reports
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    • v.39 no.6
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    • pp.717-721
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    • 2006
  • In vitro analyses of type I signal peptidase activities require protein precursors as substrates. Usually, these pre-proteins are expressed in vitro and cleavage of the signal sequence is followed by SDS polyacrylamide gel electrophoresis coupled with autoradiography. Radioactive amino acids have to be incorporated in the expressed protein, since the amount of the in vitro expressed protein is usually very low and processing of the signal peptide cannot be followed by SDS polyacrylamide gel electrophoresis alone. Here we describe a rapid and simple method to express large amounts of a protein precursor in E. coli. We have analyzed the effect of ionophors as well as of azide on the accumulation of expressed protein precursors. Azide blocks the function of SecA and the ionophors dissipate the electrochemical gradient across the cytoplasmic membrane of E. coli. Addition of azide ions resulted in the formation of inclusion bodies, highly enriched with pre-apo-plastocyanine. Plastocyanine is a soluble copper protein, which can be found in the periplasmic space of cyanobacteria as well as in the thylakoid lumen of cyanobacteria and chloroplasts, and the pre-protein contains a cleavable signal sequence at its N-terminus. After purification of cyanobacterial pre-apo-plastocyanine, its signal sequence can be cleaved off by the E. coli signal peptidase, and protein processing was followed on Coomassie stained SDS polyacrylamide gels. We are optimistic that the presented method can be further developed and applied.

Antioxidative Compounds in Aerial Parts of Potentilla fragarioides (양지꽃(Potentilla fragarioides) 지상부의 항산화물질)

  • Choi, Yong-Hwa;Kim, Myong-Jo;Lee, Haeng-Soon;Yun, Bong-Sik;Hu, Changxu;Kwak, Sang-Soo
    • Korean Journal of Pharmacognosy
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    • v.29 no.2
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    • pp.79-85
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    • 1998
  • Six antioxidative compounds in the aerial parts of Potentilla fragarioides were isolated by a bioassay guided purification using a DPPH free radical. They were identified as (+)-catechin, isoquercitrin, quercitrin, $quercetin-3-O-{\beta}-D-glucopyranosyl-{\beta}-D-xylopyranoside$, caffeic acid, and 4-O-caffeoyl-L-threonic acid on the basis of $^{1}H$ and $^{13}C-NMR$ and MS data. The DPPH radical scavenging activity of five compounds $(RC_{50}:\;7.5{\sim}10.5\;{\mu}g)$ except for quercitrin $(16\;{\mu}g)$ was more effective than those of ${\alpha}-tocopherol$ $(12\;{\mu}g)$ and BHA $(14\;{\mu}g)$.

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High-yield Expression and Characterization of Syndecan-4 Extracellular, Transmembrane and Cytoplasmic Domains

  • Choi, Sung-Sub;Kim, Ji-Sun;Song, Jooyoung;Kim, Yongae
    • Bulletin of the Korean Chemical Society
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    • v.34 no.4
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    • pp.1120-1126
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    • 2013
  • The syndecan family consists of four transmembrane heparan sulfate proteoglycans present in most cell types and each syndecan shares a common structure containing a heparan sulfate modified extracellular domain, a single transmembrane domain and a C-terminal cytoplasmic domain. To get a better understanding of the mechanism and function of syndecan-4 which is one of the syndecan family, it is crucial to investigate its three-dimensional structure. Unfortunately, it is difficult to prepare the peptide because it is membrane-bound protein that transverses the lipid bilayer of the cell membrane. Here, we optimize the expression, purification, and characterization of transmembrane, cytoplasmic and short extracellular domains of syndecan4 (syndecan-4 eTC). Syndecan-4 eTC was successfully obtained with high purity and yield from the M9 medium. The structural information of syndecan-4 eTC was investigated by MALDI-TOF mass (MS) spectrometry, circular dichroism (CD) spectroscopy, and nuclear magnetic resonance (NMR) spectroscopy. It was confirmed that syndecan-4 eTC had an ${\alpha}$-helical multimeric structure like transmembrane domain of syndecan-4 (syndecan-4 TM) in membrane environments.