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  • Title/Summary/Keyword: Outer membrane

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The Distribution of ATPase and Porin in the Bovine Heart Mitochondrial Cristae (소(牛) 심근 미토콘드리아의 ATPase와 porin의 분포)

  • Kim, Tae-Keun;Min, Byoung-Hoon;Kim, Soo-Jin
    • Applied Microscopy
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    • v.40 no.4
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    • pp.261-266
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    • 2010
  • ATP is the energy source synthesized at the electron transferase that consist of complex I, II, III, IV and V in mitochondrial cristae. The complex V functions as ATPase which composed of sub-complex F0 and F1. Porin or VDAC (voltagedependent anion-selective channel), is a family of small pore-forming proteins of the mitochondrial outer membrane, and play important roles in the regulated flux of anion, proton and metabolites between the cytosolic and mitochondrial compartments. The channel allows the diffusion of negatively charged solutes such as succinate, malate, and ATP in the fully open state, but of positively charged ions in subconducting state. In this study, in order to investigate the relationship of the function and localization between porin and ATPase we observed the distribution of porin and ATPase in the mitochondria of the bovine heart. Monoclonal antibodies against porin and ATPase β-subunit were used to detect porin and ATPase using light microscope with immunohistochemistry and immunofluorescence, and using electron microscope with immunogold-labeling. ATPase were stained in longitudinal section region in cardiac muscle, porin were stained in longitudinal section region in cardiac muscle. We viewed more specific pattern of localization and distribution of these proteins using immunofluorescence method. There were some region which were labeled with porin or ATPase respectively, and others which were labeled both proteins in cardiac muscle. The electron microscope results showed that immunogold labeled porin were labeled locally at mitochondrial outer membrane and ATPase were labeled evenly at mitochondrial cristae. But ATPase was not labeled at mitochondria cristae. These results confirmed the subcellular localizations of porin and ATPase in mitochondrial outer membrane and cristae. Also, we assumed that ATP synthesis always does not activation in all mitochondria exist in the bovine cardiac muscle.

Comparative serological analysis of outer membrane proteins extracted from Brucella abortus Korean isolates and 1119-3 strains (Brucella abortus 국내 분리주의 세포외막 단백질 분석 및 혈청학적 비교)

  • Cha, Seung-Bin;Kang, Mi-Lan;Lee, Won-Jung;Shin, Min-Kyoung;Cho, Dong-Hee;Jung, Suk-Chan;Yoo, Han-Sang
    • Korean Journal of Veterinary Research
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    • v.48 no.4
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    • pp.431-440
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    • 2008
  • Brucellosis is one of the most important zoonosis in worldwide. As one of the control measures, attempts have been made to develop new diagnostic methods using filed isolates as a national policy in many countries. Currently, bovine brucellosis in Korea have been received attention in both public health and economical aspects due to sudden increase of outbreak. Based on the situation, we compared standard strain (B. abortus 1119-3) with field isolates to reveal the differences among them. Biological and biochemical charateristics, antibiotic resistance profiles, outer membrane proteins (OMPs) and lipopolysaccharide analysis of the strains were included in this study. For the diagnostic purpose, an attempt was made to find out a novel antigen from the Korean isolates by serological analysis. There were differences about 55 kDa, 36-38 kDa and 20 kDa in analysis of OMPs by SDS-PAGE and Western blot with positive sera ( 1:400 in SAT titer). Also, a serological diagnostic method, ELISA was conducted using OMPs of the strains as novel antigen. Relationships between O.D. and SAT titer were analyzed using field sera showing different SAT titer. High correlation coefficient was observed between SAT titer and ELISA. Results from this study suggested that a new diagnsotic method should be developed using their own field isolates in each country.

Escherichia coli-Derived Outer Membrane Vesicles Deliver Galactose-1-Phosphate Uridyltransferase and Yield Partial Protection against Actinobacillus pleuropneumoniae in Mice

  • Quan, Keji;Zhu, Zhuang;Cao, Sanjie;Zhang, Fei;Miao, Chang;Wen, Xintian;Huang, Xiaobo;Wen, Yiping;Wu, Rui;Yan, Qigui;Huang, Yong;Ma, Xiaoping;Han, Xinfeng;Zhao, Qin
    • Journal of Microbiology and Biotechnology
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    • v.28 no.12
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    • pp.2095-2105
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    • 2018
  • In our previous studies, we have identified several in vivo-induced antigens and evaluated their potential as subunit vaccine candidates in a murine model, in which the recombinant protein GalT showed the most potent immunogenicity and immunoprotective efficacy against Actinobacillus pleuropneumoniae. To exploit a more efficient way of delivering GalT proteins, in this study, we employed the widely studied E. coli outer membrane vesicles (OMVs) as a platform to deliver GalT protein and performed the vaccine trial using the recombinant GalT-OMVs in the murine model. Results revealed that GalT-OMVs could elicit a highly-specific, IgG antibody titer that was comparable with the adjuvant GalT group. Significantly higher lymphocyte proliferation and cytokines secretion levels were observed in the GalT-OMVs group. 87.5% and 50% of mice were protected from a lethal dose challenge using A. pleuropneumoniae in active or passive immunization, respectively. Histopathologic and immunohistochemical analyses showed remarkably reduced pathological changes and infiltration of neutrophils in the lungs of mice immunized with GalT-OMVs after the challenge. Taken together, these findings confirm that OMVs can be used as a platform to deliver GalT protein and enhance its immunogenicity to induce both humoral and cellular immune responses in mice.

Purification and Characterization of Mitochondrial Mg2+-Independent Sphingomyelinase from Rat Brain

  • Jong Min Choi;Yongwei Piao;Kyong Hoon Ahn;Seok Kyun Kim;Jong Hoon Won;Jae Hong Lee;Ji Min Jang;In Chul Shin;Zhicheng Fu;Sung Yun Jung;Eui Man Jeong;Dae Kyong Kim
    • Molecules and Cells
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    • v.46 no.9
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    • pp.545-557
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    • 2023
  • Sphingomyelinase (SMase) catalyzes ceramide production from sphingomyelin. Ceramides are critical in cellular responses such as apoptosis. They enhance mitochondrial outer membrane permeabilization (MOMP) through self-assembly in the mitochondrial outer membrane to form channels that release cytochrome c from intermembrane space (IMS) into the cytosol, triggering caspase-9 activation. However, the SMase involved in MOMP is yet to be identified. Here, we identified a mitochondrial Mg2+-independent SMase (mt-iSMase) from rat brain, which was purified 6,130-fold using a Percoll gradient, pulled down with biotinylated sphingomyelin, and subjected to Mono Q anion exchange. A single peak of mt-iSMase activity was eluted at a molecular mass of approximately 65 kDa using Superose 6 gel filtration. The purified enzyme showed optimal activity at pH of 6.5 and was inhibited by dithiothreitol and Mg2+, Mn2+, Ni2+, Cu2+, Zn2+, Fe2+, and Fe3+ ions. It was also inhibited by GW4869, which is a non-competitive inhibitor of Mg2+-dependent neutral SMase 2 (encoded by SMPD3), that protects against cytochrome c release-mediated cell death. Subfractionation experiments showed that mt-iSMase localizes in the IMS of the mitochondria, implying that mt-iSMase may play a critical role in generating ceramides for MOMP, cytochrome c release, and apoptosis. These data suggest that the purified enzyme in this study is a novel SMase.

Intranasal Vaccination with Outer-Membrane Protein of Orientia tsutsugamushi induces Protective Immunity Against Scrub Typhus

  • Sung-Moo Park;Min Jeong Gu;Young-Jun Ju;In Su Cheon;Kyu-Jam Hwang;Byoungchul Gill;Byoung-Shik Shim;Hang-Jin Jeong;Young Min Son;Sangho Choi;Woonhee Jeung;Seung Hyun Han;Hyuk Chu;Cheol-Heui Yun
    • IMMUNE NETWORK
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    • v.21 no.2
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    • pp.14.1-14.17
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    • 2021
  • Scrub typhus develops after the individual is bitten by a trombiculid mite infected with Orientia tsutsugamushi. Since it has been reported that pneumonia is frequently observed in patients with scrub typhus, we investigated whether intranasal (i.n.) vaccination with the outer membrane protein of O. tsutsugamushi (OMPOT) would induce a protective immunity against O. tsutsugamushi infection. It was particular interest that when mice were infected with O. tsutsugamushi, the bacteria disseminated into the lungs, causing pneumonia. The i.n. vaccination with OMPOT induced IgG responses in serum and bronchoalveolar lavage (BAL) fluid. The anti-O. tsutsugamushi IgA Abs in BAL fluid after the vaccination showed a high correlation of the protection against O. tsutsugamushi. The vaccination induced strong Ag-specific Th1 and Th17 responses in the both spleen and lungs. In conclusion, the current study demonstrated that i.n. vaccination with OMPOT elicited protective immunity against scrub typhus in mouse with O. tsutsugamushi infection causing subsequent pneumonia.

Cell Structures of Spermatogenesis of Rainbow Trout, Oncorhynchus mykiss in Reproductive Cycles (번식주기(繁殖週期)에 따른 무지개 송어(松魚) 정자형성시(精子形成時) 세포구조(細胞構造)의 변화(變化))

  • Yoon, Jong-Man;Kim, Gye-Woong;Park, Chung-Kil;Roh, Soon-Chang
    • Applied Microscopy
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    • v.24 no.3
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    • pp.55-66
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    • 1994
  • This study was carried out to investigate the histological changes of sperm cells in testis, obtained from 100 of 3-year-old male rainbow trout (Oncorhynchus mykiss) collected and analysed from March in 1992 to February in 1993. Especially, the ultrastructural changes of spermatogonia, primary and secondary spermatocytes, spermatids, and spermatozoa were examined to describe the reproductive cycles of this species. The results obtained in this study were as follows: The ultrastructures of the gonadotrophs largely parallel the cyclical changes in the testes. Each nest of cells belongs to one spermatogenetic stage, although nests at different stages can be found within the one lobule. At first keterochromatin is dispersed and then is condensed. In mature gamete, the nucleus is dense and homogeneous. The nuclear membrane appeared at the beginning of differentiation. In spermatogonia, Sertoli cells are located at the periphery of their cytoplasm. In the primary spermatocytes, the small mitochondria are abundant over the outer cytoplasm. During cell differentiation, the cytoplasm decreases and the nucleus increases. In spermatids, the protein masses moved towards the posterior part of the nucleus. In late spermatids, the two large mitochondria are located over the cytoplasm. In spermatozoa, two spheroidal mitochondria (about 145nm long) are situated in parallel between the nucleus and the axoneme. Spermatozoa mitochondria are assembled into an organized sheath surrounding the outer dense fibres and axoneme of the flagellar midpiece. The two centrioles are quite separate and the central pair and sheath complex of the flagellum is inserted into the base of the distal centriole.

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Redescription of Canthocamptus mirabilis Sterba (Copepoda, Harpacticoida), Based on the Topotypic Material from China (중국의 모식산지에서 채집한 갈고리노벌레류 Canthocamptus mirabilis Sterba의 재기재)

  • Chang, Cheon-Young
    • Animal Systematics, Evolution and Diversity
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    • v.17 no.1
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    • pp.1-11
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    • 2001
  • Conthocamptus mirabilis Sterba is redescribed on the basis of the material newly collected from the type locality near Bekijing, China, with an evaluation of the variability of several characters. C. mirabilis possesses the plesiomorphic characters of the round and narrow hyaline membrane and the unmodified outer terminal furcal setae, while it also bears the apomorphic ones of the male-type caudal rami in females and the modified outer terminal setae of male leg 4 exopod. the peculiar dorsal keel ornamented with spinule array on the female's caudal rami is the most significant feature differentiating this species from its relatives of the mirabilis group, which turned out to be a species group of closely related and largely allopatric or parapatric species. Affinities with the allies of the species group are discussed, and the morphological discrepancies differentiating the Japanese population are enumerated.

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Taxonomy on Canthocamptus semicirculus and C. coreensis n. sp.(Harpacticoida, Canthocarnptidae), with a Key to the C. mirabilis Species Group from South Korea (딱정 장수노벌레속(갈고리노벌레목, 딱정장수노벌레과) mirabilis 종군에 속하는 1신종 1기록종의 분류학적 연구)

  • Chang, Cheon-Young
    • Animal Systematics, Evolution and Diversity
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    • v.18 no.2
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    • pp.233-244
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    • 2002
  • As one of the serial studies on the taxonomy of Conthocamptus mirabilis species group in South Korea, C. semicirculus Kikuchi, widely distributed in the southern part of the Far East, and C. coreensis n. sp. from the middle west of South Korea are recorded. Intraspecific variability of some important characters like outer caudal setae and the spinous process of male leg 3 exopod was examined in Korean population of C. semicirculus. Canthocamptus coreensis n. sp. possesses the plesiomorphic characters of the round and narrow hyaline membrane of anal operculum and lacking the sexual reverse transformation in female caudal rami, while it also evolves the apomorphic ones of the modified outer apical setae of male leg 4 exopod and the spinous process of male leg 3 exopod. A key to the five species of the species group known from Korea is prepared.

Analysis of outer mombrane proteins of Brucella abortus using two dimensional polyacrylamide gel electrophoresis (2차원 전기영동법을 이용한 Brucella abortus 세포외막 특이단백질의 분석)

  • Kim, Byung-su;Kim, Sun-hee;Kim, Jong-suk;Baek, Byeong-kirl
    • Korean Journal of Veterinary Research
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    • v.38 no.2
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    • pp.328-335
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    • 1998
  • Outer membrane proteins(OMPs) of Brucella abortus 1119-3 strain were extracted by Triton X-100 treatment, and fractionated by DEAE-cellulose column chromatography and Sephacryl S-300 column chromatography. The antigenic proteins in these fractions were identified by Western blot analysis. In Western blot analysis, a single band(38kDa) was observed in the DEAE fractions from 36th fraction to 38th fraction against sera of cattle infected with B abortus. And other fractions have several bands. However, the Sephacryl S-300 fractions exhibited a total of 3 peaks of proteins with a broad range from about 30 to 116kDa. In order to characterize further, the extracted OMPs and the DEAE fractions were analyzed by two dimensional polyacrylamide gel electrophoresis(2-DE) and Western blot using serum from naturally infected cattle with Brucella spp. The 2-DE immunoblots of DEAE fraction showed immunoreactive spots more than twenty two. The major protein spots have ranging from about 32 to 47kDa. The pI values of the spots were detected from pH 4.7 to 5.4. Among the major protein spots, the 38kDa protein which is a specific antigen, located at the point of approximately a pI 4.8.

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Physics on cancer and its curing

  • Oh, Hung-Kuk
    • Proceedings of the Korean Society for Emotion and Sensibility Conference
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    • 2000.11a
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    • pp.91-97
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    • 2000
  • The conventional model did not take momentum conservation into consideration when the electron absorbs and emits the photons. II-ray provides momentum conservations on any directions of the entering photons, and also the electrons have radial momentum conservations and fully elastic bouncing between two atoms, in the new atom model. Conventional atom model must be criticized on the following four points. (1) Natural motions between positive and negative entities are not circular motions but linear going and returning ones, for examples sexual motion, tidal motion, day and night etc. Because the radius of hydrogen atom's electron orbit is the order of 1011 m and the radia of the nucleons in the nucleus are the order of 10l4m and then the converging n-gamma rays to the nucleus have so great circular momentum, the electron can not have a circular motion. We can say without doubt that any elementary mass particle can have only linear motion because of the n-rays' hindrances, near the nucleus. (2) Potential energy generation was neglected when electron changes its orbit from outer one to inner one. The h v is the kinetic energy of the photo-electron. The total energy difference between orbits comprises kinetic and potential energies. (3) The structure of the space must be taken into consideration because the properties of the electron do not change during the transition from outer orbit to inner one even though it produces photon. (4) Total energy conservation law applies to the energy flow between mind and matter because we daily experiences a interconnection between mind and body. An understanding of the mechanisms responsible for the control of normal proliferation and differentiation of the various cell types which make up the human body will undoubtedly allow a greater insight into the abnormal growth of cells, A large body of biochemical evidence was eventually used to generate a receptor model with an external ligand binding domain linked through a single trans-membrane domain to the cytoplasmic tyrosine kinase and autophosphory-lation domains. The ligand induced conformational change in the external domain generates either a push-pull or rotational signal which is transduced from the outside to the inside of cell.l.ell.

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