• Title/Summary/Keyword: Chain complexes

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Fine Structure and Immunogoldlabeling of Crystalline Inclusion Bodies in Mitochondria (미토콘드리아내 결정함유물의 미세구조 및 면액황금표식법)

  • ;;R.A. Capaldi
    • The Korean Journal of Zoology
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    • v.31 no.1
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    • pp.62-70
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    • 1988
  • The distribution of respiratory chain complexes in beef heart and human muscle mitochondria has been explored by immunoeledron microscopy with antibodies made against beef heart mltochondriai proteins in conjundion with protein A cofloidai gold (l2nm particles). The antibodies used were made against NADH-conezyme Q reductase(complex I), ubiquinol-cytochrome-c-oxldoreductase (complex III) and cytochrome-c-oxidase(complex IV). Labeling of bed heart tissue with any of these antihodies gave gold particles randomly distributed along the mitochondrial inner membrane. The labeling of muscle tIssue mitochondria from a patient with a mitochondrial myopathy localized by biochemical analysis to complex III was quantitated and compared with the labeling of human control muscle tissue mitochondria. Four kinds of morphological changes in the mitochondrial fine strudure in the myopathy patient tissue have been found; paracrystalline inclusions consistIng of densely packed multi- lamellar structures, globular crystalline inclusions with high electron density, multilamellar strudure inclusion body(compadly and irregularly arranged concentric whirl shaped cristae)and golbular cyrstalilne inclusions located in the center of the whirl shaped cristae. Compex I and cytochrome-c-oxldase antihodies reacted to the same level in the mitochondria containing the crystalline inclusions and control mitochondria. Antibodies to complex III reacted very poorly to the mitochondria containing the crystalline Inclusions but strongly to control mitchondria. The globular crystalline inclusions in the mitochondria are not reacted antibodies to respiratory chain complexes.

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Agronomic characteristics of stay-green mutant derived from an early-maturing rice variety 'Pyeongwon'

  • Won, Yong-Jae;Ji, Hyeon-So;Ahn, Eok-Keun;Lee, Jeong-Heui;Jung, Kuk-Hyun;Lee, Sang-Bok;Hong, Ha-Cheol;Hyun, Ung-Jo;Ha, Woon-Goo;Kim, Myeong-Ki;Kim, Byeong-Ju
    • Proceedings of the Korean Society of Crop Science Conference
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    • 2017.06a
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    • pp.72-72
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    • 2017
  • We found a new stay-green mutant from 'Pyeongwon' which is an early-maturing rice variety in Korea. The mutant showed green leaves after grain ripening period and it maintained higher SPAD value than wild type rice plant and original variety 'Pyeongwon'. The stay-green trait in rice, three genes have been identified up to date. The non-yellow coloring1 (NYC1) gene encodes a chloroplast-localized short-chain dehydrogenase/reductase (SDR) with three transmembrane domains. The non-yellow coloring3 (NYC3) gene encodes a plastid-localizing alpha/beta hydrolase-fold family protein with an esterase/lipase motif. The Sgr gene encodes a novel chloroplast protein and regulates the destabilization of the light-harvesting chlorophyll binding protein (LHCP) complexes of the thylakoid membranes, which is a prerequisite event for the degradation of chlorophylls and LHCPs during senescence. After sequencing the PCR products, we found a single nucleotide variation($A{\rightarrow}T$) in the NYC1 gene, which changes the amino acid lysine to methionine. The NYC1 gene encodes a short-chain dehydrogenase/reductase(SDR) protein. And we confirmed the co-segregation between SNP and stay-green trait from genotyping the progenies of the mutant.

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CD7-Specific Single Chain Antibody Mediated Delivery of siRNA to T Cells Inhibits HIV Replication in a Humanized Mouse Model

  • Ban, Hong-Seok;Kumar, Priti;Kim, Na-Hyun;Choi, Chang-Son;Shankar, Premlata;Lee, Sang-Kyung
    • Proceedings of the Microbiological Society of Korea Conference
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    • 2008.05a
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    • pp.62-64
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    • 2008
  • A major hurdle to the development of RNA interference as therapy for HIV infection is the delivery of siRNA to T lymphocytes which are difficult cells to transfect even in vitro. We have employed a single chain antibody to the pan T cell surface antigen CD7 was conjugated to an oligo-9-arginine peptide (scFvCD7-9R) for T cell-specific siRNA delivery in NOD/SCIDIL2${\gamma}$-/- mice reconstituted with human peripheral blood lymphocytes (Hu-PBL). Using a novel delivery, we first show that scFvCD7-9R efficiently delivered CD4 siRNA into human T cells in vitro. In vivo administration to Hu-PBL mice resulted in reduced levels of surface CD4 expression on T cells. Mice infected with HIV-1 and treated on a weekly basis with scFvCD7-9R-siRNA complexes targeting a combination of viral genes and the host coreceptor molecule CCR5 successfully maintained CD4/CD3 T cell ratios up to 4 weeks after infection in contrast to control mice that displayed a marked reduction in CD4 T cell numbers. p24 antigen levels were undetectable in 3 of the 4 protected mice. scFvCD7-9R/antiviral siRNA treatment also helped maintain CD4 T cell numbers with reduced plasma viral loads in Hu-PBL mice reconstituted with PBMC from donors seropositive for HIV, indicating that this method can contain viral replication even in established HIV infections. Our results show that scFvCD7-9R could be further developed as a potential therapeutic for HIV-1 infection.

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Effect of Salt Concentration on the Glass Transition Temperature and Ionic Conductivity of Poly(ethylene glycol)-Polyurethane/$LiClO_4$ Complexes

  • Huh, Pil-Ho;Park, Myung-Geun;Jo, Nam-Ju;Lee, Jin-Kook;Lee, Jang-Oo;Wongkang Yang
    • Macromolecular Research
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    • v.12 no.4
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    • pp.422-426
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    • 2004
  • Solid polymer electrolytes based on poly(ethylene glycol)-polyurethane (PEG-PU) complexed with LiClO$_4$ salt have been prepared by the solvent casting method. A PEG-PU material (PEG:4,4'-diphenylmethane diisocyanate: l,4-butanediol = 1:2:1) was synthesized through a typical two-step condensation reaction. We investigated the effects of the salt concentration on the ionic conductivity ($\sigma$) and the glass transition temperature (T$_{g}$ ) of the complex electrolytes by using alternating current impedance spectroscopy, differential scanning calorimetry, and dynamic mechanical thermal analysis. The measured values of both $\sigma$ and T$_{g}$ exhibited similar tendencies in that they had maxima within the range studied, probably because of two opposite effects, i.e., the increased number of carrier ions and the decreased chain mobility (or increased T$_{g}$ ) caused by the increase in the salt concentration. The highest conductivity, on the order of 2.43 ${\times}$ 10$^{6}$ S$cm^{-1}$ /, was obtained at an [O]/[Li$^{+}$] ratio of ca. 16 (0.92 ㏖ salt per kg of matrix polymer).

Ferritin, an Iron Storage Protein, Associates with Kinesin 1 through the Cargo-binding Region of Kinesin Heavy Chains (KHCs) (철 저장 단백질 ferritin과 kinesin 1 결합 규명)

  • Jang, Won Hee;Jeong, Young Joo;Lee, Won Hee;Kim, Mooseong;Kim, Sang-Jin;Urm, Sang-Hwa;Moon, Il Soo;Seog, Dae-Hyun
    • Journal of Life Science
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    • v.26 no.6
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    • pp.698-704
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    • 2016
  • The intracellular transport of organelles and protein complexes is mediated by kinesin superfamily proteins (KIFs). The first kinesin, kinesin 1, was identified as a molecular motor protein that moves various organelles and protein complexes along the microtubule rails in cells. Kinesin 1 is a tetramer of two heavy chains (KHCs, also called KIF5s) and two kinesin light chains (KLCs). KIF5s interact with many different proteins through their tail region, but their binding proteins have not yet been fully identified. To identify the interaction proteins for KIF5A, we performed yeast two-hybrid screening and found a specific interaction with ferritin heavy chain (Frt-h), which has a role in iron storage and detoxification. Frt-h bound to the amino acid residues between 800 and 940 of KIF5A and to other KIF5s in the yeast two-hybrid assay. The coiled-coil domain of Frt-h is essential for interaction with KIF5A. In addition, ferritin light chain (Frt-l) interacted with KIF5s in the yeast two-hybrid assay. In addition, these proteins showed specific interactions in the glutathione S-transferase (GST) pull-down assay. An antibody to KHC specifically co-immunoprecipitated Frt-h and Frt-l from mouse brain extracts. These results suggest the kinesin 1 motor protein may transport the ferritin complex in cells.

SYNTHESIS AND CHARACTERIZATION OF THE POLYCATIONS FOR THE PREPAEATION OF POLYION COMPLEX MEMBRANS

  • Jegal, J.G.;park, Y.I.;Kim, J.H.;Lee, K.H.
    • Proceedings of the Membrane Society of Korea Conference
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    • 1993.10a
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    • pp.58-58
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    • 1993
  • Pervaporation has become important as a method to separate liquid mixtures, sepecially azeotropic and close boiling-point mixtures. Especially, water-alcohol separations have been carried out a lot because of the practical interests in the industries. However, outstanding membranes with high selectivity and high permeability have not been available in common use yet. In order to separate selectively out the water mixed with alcohols with an aim of the purification of the alcohols, a membrane has tO have excellent affinity to water. Among the hydrophilic polymers, polyacrylic acid and poly(vinyl alcohol) are used widely. In recent years, ionomers and polyion complexes, better hydrophitic materials, start to be used. The polyion complex membranes, consisting of polyacrylic acid (PAA) and polycation, showed excellent permeation rates and selectivities. It was known that among the polycations, ioneries, which have quaternary ammonium groups in the backbone chain, were more effective in giving membranes of higher permselectivities. On this base, syntheses and characterizations of the polycations, with different chemical structures from the published ones, for the polyion complex membrane formation were studied in this paper.

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Localization of Autophagosome in Porcine Follicular Cumulus-oocyte Complex

  • Lee, Seunghoon;Kim, Dong-Hoon;Im, Gi-Sun;Ock, Sun-A;Ullah, Imran;Hur, Tai-Young
    • Journal of Embryo Transfer
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    • v.32 no.3
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    • pp.105-109
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    • 2017
  • Autophagy is an intracellular degradation and recycling system. Oocyte maturation is dynamic process, in which various proteins should be synthesized and degraded. In our previous study, we reported the loci of autophagosome and dynamics of autophagic activity in porcine oocytes during in vitro maturation. In this study, we verified loci of autophagosome in porcine follicular cumulus-oocyte complex by detection of microtubule-associated protein 1A/1B-light chain 3 (LC3) which is the reliable marker of autophagosome. Porcine ovary including various sizes of follicles was fixed within 1 hour after collection from slaughterhouse. After fixation, immunohistochemistry was conducted on sliced ovary tissue containing various sizes of follicles by using LC3 antibody. As a result, LC3 signal was clearly detected in both cumulus and oocytes of various sizes of follicles. We also found ring shaped signal which represent autophagosome near oocyte membrane. Most of the signals in oocytes were localized nearby cellular membrane while evenly dispersed in cumulus cells. Therefore, this result suggests that autophagy occurs in porcine COCs (cumulus-oocyte complexes) at follicular stage.

Precipitation, Resolubilization and Luminescent Properties of Tris (2,2$^\prime$-diimine)Ruthenium(II) Complexes in Premicellar Anionic Surfactant Solutions

  • Park, Joon-Woo;Kim, Sung-Hee
    • Bulletin of the Korean Chemical Society
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    • v.9 no.5
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    • pp.317-322
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    • 1988
  • Premicellar precipitation, resolubilization and luminescing behaviors of $RuL_3^{2+}$ (L = bpy, phen, $Me_2bpy$) in aqueous alkylsulfate and sulfonate solutions were studied. Addition of the anionic surfactants to $RuL_3\;^{2+}$ solutions caused initial precipitation which was redissolved by further addition of the surfactants. The apparent solubility products $K_{sp}$'s of the precipitates were evaluated assuming 1:2 salt formation. The values were smaller as the ligand is more hydrophobic and the length of hydrocarbon chain of the surfactant is longer. The $K_{sp}$ values for L = bpy were constant over wide surfactant concentration range. However, those for L = $Me_2bpy$ and also for phen, but to less extent, increased with the surfactant concentration. The resolubilization of 1:2 salts was followed by red-shift of emission band and extensive emission quenching above critical concentration of the surfactants. The critical concentration was lower for more hydrophobic surfactant. For L = $Me_2bpy$, the blue-shifted emission band with enhanced emission intensity was observed in intermediate surfactant concentration region. The high ionic strength of media prevented the precipitate formation, but facilitated the red-shift of the emission bands. The results support that the precipitate is dissolved by accretion of surfactant anions to the salts to form water-soluble surfactant-rich $RuL_3$-surfactant anionic species. These species appeared to aggregate cooperatively to produce large clusters which exhibited the red-shifted emission.

Intermacromolecular Complex Formation between Helix Strilctilral Polypeptides through Hydrogen Bonding (수소 결합을 통한 Helix 폴리 펩타이드사이의 복합체 형성)

  • 조병기;김창규
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.18 no.1
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    • pp.99-132
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    • 1992
  • Polypeptide has been used broadly as an active ingredient in cosmetics We thought it is very important to investigate the adsorption behavior of polypeptide in order to pre-estimate the effect of these polypeptides. For the study of polypeptide adsorption, we have investigated complex formation of basic homopolypeptides, poly(L-proline) Form I [PLP(I)], Form II [PLP(II)] and poly(4-hydroxy-L-proline) (PHLP) with acidic homopolypeptides, poly(L-glutamic acid) (PLGA), poly(D-glutamic acid) (PDGA) and poly(L-aspartic acid) (PLAA) through hydrogen bonding in a hydroalcoholic medium with viscometer, 1ight scatter, pH meter and circular dicroism (CD). The polypeptides used in this study have helical structure in some conditions. The result exhibited that al 1 the complexes were formed as the composition of basic/acidic homopolypeptide : L:2 irrespective of the complex systems used. A more favorable complex is formed in the PLP(II)-PLGA system than PHLP-PLGA because PLP(II) has a more flexible helical conformation, whereas PHLP has a more rigid helical conformation. The right-handed helix PLGA formed the complex favorably and quickly with the left-handed helix PLP(II), whereas the left-handed helix PDGA formed the complex favorably with the right-handed helix PLP(I). The effect of side chain of the acidic homopolypeptides on the complexation was also studied. The result showed that more favorable condition for the complexation was PLGA-PLP(II) system which has longer side chain at acidic homopolypeptide than PLAA - PLP(II). All the above facts were well supported by CD measurement for the complex systems. By the CD spectra for the complexes we could deduce the conformational change of each homopolypeptide in the complexes On the basis of the above results, we performed the adsorption test of PLP(I, II) and PHLP on the hair having a left-handed helix. The adsorption amount of each polypeptide was analyzed by HPLC. The result showed that PLP(I) was adsorbed more than PLP(II), PLP(II) was adsorbed more than PHLP on the hair. On adsorbing polypeptides having a helical structure on the hair through hydrogen bonding, it could be concluded that the helical polypeptides having the opposite directional structure to the hair are adsorbed more than those having the same directional structure with the hair and also the polypeptides having a flexible conformation are adsorbed more than those having a rigid conformation.

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Rab Effector EHBP1L1 Associates with the Tetratricopeptide Repeat Domain of Kinesin Light Chain 1 (Kinesin Light Chain 1 (KLC1)의 Tetratricopeptide Repeat (TPR) 도메인과 Rab effector, EHBP1L1의 결합)

  • Jeong, Young Joo;Park, Sung Woo;Kim, Sang-Jin;Kim, Mooseong;Urm, Sang-Hwa;Lee, Jung Goo;Seog, Dae-Hyun
    • Journal of Life Science
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    • v.30 no.1
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    • pp.10-17
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    • 2020
  • Kinesin-1 is microtubule-dependent plus-end direct molecular motor protein essential for intracellular transport. It is a member of the kinesin superfamily proteins (KIFs) which transport cargo, including organelles, vesicles, neurotransmitter receptors, cell-signaling molecules, and protein complexes through interaction between its light chain subunit and the cargo. Kinesin light chain 1 (KLC1) is a non-motor subunit that associates with the kinesin heavy chain (KHC). Although KLC1 interacts with many different adaptor proteins and scaffolding proteins, its binding proteins have not yet been fully identified. We used the yeast two-hybrid assay to identify proteins that interact with the tetratricopeptide repeat (TPR) domain of KLC1, and found an interaction between KLC1 and EH domain-binding protein 1 like 1 (EHBP1L1). EHBP1L1 bound to the region containing all six TPR repeats of KLC1 and did not interact with KIF5B (a motor protein of kinesin 1) or KIF3A (a motor protein of kinesin 2) in the yeast two-hybrid assay. The carboxyl-terminus of the coiled-coil domain of EHBP1L1 is essential for interaction with KLC1. However, another EHBP1L1 isoform, EHBP1, did not interact with KLC1 in the yeast two-hybrid assay. KLC1 interacted with GST-EHBP1L1 and its coiled-coil domain but not with GST only. When co-expressed in HEK-293T cells, EHBP1L1 co-localized with KLC1 and co-immunoprecipitated with KLC1 and KIF5B but not KIF3A. These results suggest that kinesin 1 motor protein may transport EHBP1L1-associated cargo in cells.