• Title/Summary/Keyword: Membrane Protein

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Structural Characterization of Hordeum vulgare L. Chloroplast by Ozone

  • Chung, Hwa-Sook;Lim, Young-Jin;Park, Kang-Eun
    • Environmental Sciences Bulletin of The Korean Environmental Sciences Society
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    • v.4 no.2
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    • pp.85-94
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    • 2000
  • The effects of ozone on chloroplast development in barley seedlings during greening was investigated based on ultrastructural changes in the chloroplasts and band pattern changes in the chloroplast thylakoid membrane proteins. In this analysis of the chloroplast thylakoid membrane thylakoid protein band pattern by SDS-PAGE, none of the 24-hour greening bands included were clearer than the control. This means that the ozone treatment produced a dealy in chloroplast development and decreased the amount of thylakoid membrane proteins. LHC II chloroplast band of developing barley seedlings treated with 0.5 and 1.0 ppm ozone during the last 4 hours of the 24-hour greening period was weaker than the other bands. This result indicates that ozone affects the LHC II protein complex of the chloroplast thylakoid membrane. When investigating the ultastructural changes in ozone-treated chloroplast, the main site affected by 0.5 ppm ozone was the chloroplast grana, thereby explaining the delayed chloroplast development during the early phase of greening. In addition, there was also a structural change in the stromal grana of the ozone treated chloroplast during the middle phase of greening. The effects of ozone on the chloroplast of barley seedlings during the last phase of 48-hour greening were more functionally inhibiting than structural changes.

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Sorting of the Human Folate Receptor in MDCK Cells

  • Kim, Chong-Ho;Park, Young-Soon;Chung, Koong-Nah;Elwood, P.C.
    • BMB Reports
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    • v.37 no.3
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    • pp.362-369
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    • 2004
  • The human folate receptor (hFR) is a glycosylphosphatidylinositol (GPI) linked plasma membrane protein that mediates delivery of folates into cells. We studied the sorting of the hFR using transfection of the hFR cDNA into MDCK cells. MDCK cells are polarized epithelial cells that preferentially sort GPI-linked proteins to their apical membrane. Unlike other GPI-tailed proteins, we found that in MDCK cells, hFR is functional on both the apical and basolateral surfaces. We verified that the same hFR cDNA that transfected into CHO cells produces the hFR protein that is GPI-linked. We also measured the hFR expression on the plasma membrane of type III paroxysmal nocturnal hemoglobinuria (PNH) human erythrocytes. PNH is a disease that is characterized by the inability of cells to express membrane proteins requiring a GPI anchor. Despite this defect, and different from other GPI-tailed proteins, we found similar levels of hFR in normal and type III PNH human erythrocytes. The results suggest the hypothesis that there may be multiple mechanisms for targeting hFR to the plasma membrane.

Adsorption Characteristic of L-tryptophan of Affinity Membrane (친화막의 L-tryptophan 흡착특성)

  • Byun, Hong-Sik;Hong, Byung-Pyo
    • Membrane Journal
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    • v.18 no.3
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    • pp.214-218
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    • 2008
  • Protain affinity membranes based on PES-BSA was preapared by the electrospinning method. The process problem caused by the electrospining was solved by using HFB having high solubility and boiling point. It was expecting that the mass production of protein affinity membrane would be possible with broad range of optimized temperature and humidity. BSA in the PES nanofiber was confirmed by the color change from colorless to violet during the biuret test. The buffer solution with DMSO showed that the amount of elution was 5 times higher than the one when the buffer solution without DMSO was used. This is due to the restriction effect of DMSO on the dissociation of L-tryptophan from BSA during the washing step.

Cloning of cDNA Encoding Putative Cellular Receptor Interacting with E2 protein of Hepatitis C Virus (C형 간염바이러스 E2 단백질에 결합하는 추정 세포수용체 cDNA의 클로닝)

  • 이성락;백재은;석대현;박세광;최인학
    • Journal of Life Science
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    • v.13 no.4
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    • pp.541-550
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    • 2003
  • E2 glycoprotein of hepatitis C virus (HCV) comprises a surface of viral particle together with E1 glycoprotein, and is thought to be involved in the attachment of HCV viral particle to receptor (s) on the permissible cells including hepatocytes, B cells, T cells, and monocytes. We constructed a phage library expressing cellular proteins of hepatocytes on the phage surface, which turned out to be 8.8${\times}$$10^5$ cfu of diversity and carried inserts in 95% of library. We screened both cDNA phage library and 12-mer peptide library to identify the cellular proteins binding to E2 protein. Some intracellular proteins including tensin and membrane band 4.1 which are involved in signal transduction of survival and cytoskeleton organization, were selected from cDNA phage library through several rounds of panning and screening. On the contrary, membrane proteins such as CCR7, CKR-L2, and insulin-like growth factor-1 receptor were identified through screening of peptide library. Phages expressing peptides corresponding to those membrane proteins were bound to E2 protein specifically as determined by neutralization of binding assay. Since it is well known that HCV can infect T cells as well as hepatocytes, we examined to see if E2 protein can bind to CCR7, a member of C-protein coupled receptor family expressed on T cells, using CCR7 transfected tells. Human CCR7 cDNA was cloned into pcDNA3.1(-) vector and transfected into human embryonic kidney cell, 293T, and expressed on the surface of the cell as shown by flow cytometer. Binding assay of E2 protein using CCR7 transfected cells indicated that E2 protein bound to CCR7 by dose-dependent mode, giving rise to the possibility that CCR7 might be a putative cellular receptor for HCV.

Antimicrobial Activity of Gluten Hydrolysate with Asp. saitoi Protease (밀 단백 효소 가수분해물의 항균활성)

  • Lee, Sang-Duk;Joo, Jeong-Hyeon;Lee, Gyu-Hee;Lee, K.T.;Oh, Man-Jin
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.32 no.5
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    • pp.745-751
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    • 2003
  • This study was carried out to investigate whether peptide produced from wheat protein by enzyme hydrolysis can be used as a natural antimicrobial agent. Antimicrobial peptide was obtained from wheat protein hydrolyzed by 7 of pretense. The produced antimicrobial peptide was purified through ultrafiltration, membrane filtration and HPLC and molecular weight and amino acid sequence of the purified antimicrobial peptide were determined. Among hydrolysate produced from wheat protein by 7 of protease, antimicrobial activity was observed for the peptide obtained from Asp. saito protease. The Asp. saito protease did produce antimicrobial hydrolysate showing the highest antimicrobial activity at reaction condition of 37$^{\circ}C$ and pH 6.0, but not at reaction condition above 5$0^{\circ}C$. Wheat protein hydrolysate was fractionated by membrane filtration and showed antimicrobial activity between molecular weight 1,000~3,000. The antimicrobial activity fraction obtained by membrane filtration was separated through HPLC and showed antimicrobial activity in the peak of retention time 31.1~31.8 min. We could convince this hydrolysate as heat-stable peptide since antimicrobial activity was maintained after treated with heat for 15 min at 121$^{\circ}C$. Molecular weight of antimicrobial peptide identified by MALDI-mass was 1,633. Amino acid sequence of antimicrobial peptide was cysteine, glycine, prolin, prolin, prolin, valine, valine, alanine, alanine and arginine.

A Study on Change of Excess Volume in Membrane of Holobacterium Holobium (Halobacterium Halobium의 Membrane에서 잉여부피 변화에 관한 연구)

  • Kim, Ki-Jun;Lee, Joo-Youb
    • Journal of the Korean Applied Science and Technology
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    • v.28 no.2
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    • pp.247-250
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    • 2011
  • The excess volumes of mixing of benzyl alcohol and halothane in vesicle and in suspensions of several lipids have been determined at $25^{\circ}C$ it using a excess volume dilatometer. The potency of general anesthetics has long been known to correlate with lipid solubility. Denaturations of the vesicle, which is a sole membrane protein in the purple membrane of Halobacteriun Halobium, were studied by absorption changes at 280 nm and fluorescence changes at 330 nm and excess volume dilatometer. The particle size analysis of viscous polymer solutions by diffusional interchange is the key step by measurement. The excess volume of mixing in chitosan was found to be negative, whereas them of purple membrane, Halobacteriun Halobium and red membrane were positive in benzyl alcohol and halothane. This result was confirmed as Miller's supposition.

Determination of Frequency of Epstein-Barr Virus in Non-Hodgkin Lymphomas Using EBV Latent Membrane Protein 1 (EBV-LMP1) Immunohistochemical Staining

  • Ishtiaq, Sheeba;Hassan, Usman;Mushtaq, Sajid;Akhtar, Noreen
    • Asian Pacific Journal of Cancer Prevention
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    • v.14 no.6
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    • pp.3963-3967
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    • 2013
  • Background: The presence of Epstein-Barr virus (EBV) in Non-Hodgkin's lymphoma can be identified by immunohistochemistry for detection of EBV latent membrane protein (LMP). The role of EBV as an etiologic agent in the development of non-Hodgkin lymphoma has been supported by detection of high levels of latent membrane protein 1 (LMP-1) expression in tumors. However, no study has been conducted in a Pakistani population up till now to determine the frequency of Epstein-Barr virus positivity. The objective of our study was to determine a value for non-Hodgkin lymphoma patients using EBV LMP-1 immunostaining in our institution. Materials and Methods: This study was carried out at the Department of Histopathology, Armed Forces Institute of Pathology (AFIP), Pakistan from December 2011 to December 2012. It was a cross sectional study. A total of 71 patients who were diagnosed with various subtypes of NHL after histological and EBV LMP-1 immunohistochemical evaluation were studied. Sampling technique was non-probability purposive. Statistical analysis was achieved using SPSS version 17.0. Mean and SD were calculated for quantitative variables like patient age. Frequencies and percentages were calculated for qualitative variables like subgroup of NHL, results outcome of IHC for EBV and gender distribution. Results: Mean age of the patients was $53.6{\pm}16$ years (Mean${\pm}$SD). A total of 50 (70.4%) were male and 21 (29.6%) were female. Some 9 (12.7%) out of 71 cases were positive for EBV-LMP-1 immunostaining, 2 (22.2%) follicular lymphoma cases, 1 (11.1%) case of T-cell lymphoblastic lymphoma, 4 (44.4%) cases of diffuse large B cell lymphomas, 1 (11.1%) mantle cell lymphoma and 1 (11.1%) angioimmunoblastic T cell lymphoma case. Conclusion: In our study, frequency of EBV in NHL is 12.7% and is mostly seen in diffuse large B cell lymphoma. This requires further evaluation to find out whether this positivity is due to co-infection or has a role in pathogenesis.

Effects of Egg Yolk Antibodies Produced in Response to Different Antigenic Fractions of E. coli O157:H7 on E. coli Suppression

  • Chae, H.S.;Singh, N.K.;Ahn, C.N.;Yoo, Y.M.;Jeong, S.G.;Ham, J.S.;Kim, D.W.
    • Asian-Australasian Journal of Animal Sciences
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    • v.19 no.11
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    • pp.1665-1670
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    • 2006
  • The objective of this research was to provide the characterization and method for producing anti-E. coli O157:H7 antibodies in egg-laying hens and to determine if the antibody can restrain the proliferation of E. coli O157:H7 in-vitro. Selected antigenic fractions (whole cell, outer membrane protein and lipopolysaccharide (LPS)) from E. coli O157:H7 were injected to hens in order to produce anti-E. coli O157:H7 antibodies. The immune response and the egg yolk antibodies of laying hens against the whole cell, outer membrane protein and LPS antigens were monitored by ELISA. The level of antibodies against whole cell antigen monitored through ELISA sharply increased after the initial immunization, and it was found to be maximum on day 49 however, the level was maintained up to day 70. Antibodies (5 mg/ml) directed against the whole cell inhibited E. coli proliferation 10-13 times more than outer membrane protein or LPS. The antibody response against the whole cell antigens appeared to have higher activity in restraining the proliferation of E. coli O157:H7 than antibody against outer membrane protein or LPS. Results reflected that increasing the IgY's in the egg yolk could prevent greater economic losses due to human and animal health from pathogenic bacteria i.e. E. coli O157:H7.