• Title/Summary/Keyword: Aplysia

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Analysis of the Effects of Overexpression of Specific Phospholipid Binding Proteins on Cellular Morphological Changes in HEK293T Cells (특정 인지질 결합 단백질의 과발현이 HEK293 세포모양에 미치는 영향 분석)

  • Jun, Yong-Woo;Lee, Jin-A;Jang, Deok-Jin
    • Journal of Life Science
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    • v.26 no.8
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    • pp.875-880
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    • 2016
  • The plasma membrane plays a crucial role in relaying signals from the outside environment to the inside of the cells. In eukaryotic cells, the inner leaflets of the plasma membrane are composed mostly of phospholipids, including phosphatidylethanolamine (PE), phosphatidylserine (PS), and phosphatidylinositides (PIs). In this study, we tried to analyze the morphological changes induced by EGFP-fused membrane binding proteins, which are targeted to the plasma membrane via specific phospholipids binding. As a result, we found that overexpression of EGFP-P4M-SidM, a specific PI4P binding protein, or EGFP alone, did not induce any morphological changes. On the other hand, overexpression of EGFP-PLCδ1(PH), which is a specific PI(4,5)P2 binding protein, EGFP-AKT1(PH) which binds to PI(3,4,5)P3, or EGFP-OSH2(PH)×2 which binds to PI4P and PI(4,5)P2, could induce the filopodia and lamilapodia formation as well as cell shrinkage. Overexpression of Lact-C2-EGFP which is a specific PS-binding probe, EGFP fused Aplysia phosphodiesterase 4 (ApPDE4) long-form (L(N20)-EGFP) which is localized to the plasma membrane via hydrophobic interaction, or EGFP fused Aplysia PDE4 short-form (S(N-UCR1-2)-EGFP) which is localized to the plasma membrane via electrostatic interaction, could induce cell shrinkage, but not filopodia or lamilapodia formation. Taken together, our data support that the different phospholipid bindings in the plasma membrane could induce different characteristic morphological changes. Thus, we can analyze, characterize, and classify the cellular morphological changes induced by the various phospholipid binding proteins.

Interaction between Invertebrate Grazers and Seaweeds in the East Coast of Korea (동해안 조식성 무척추동물과 해조류 간 상호작용)

  • Yoo, J.W.;Kim, H.J.;Lee, H.J.;Lee, C.G.;Kim, C.S.;Hong, J.S.;Hong, J.P.;Kim, D.S.
    • The Sea:JOURNAL OF THE KOREAN SOCIETY OF OCEANOGRAPHY
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    • v.12 no.3
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    • pp.125-132
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    • 2007
  • We estimated the distribution of predator-prey interaction strengths for 12 species of herbivores (including amphipods, isopods, gastropods, and sea urchins) and made a regression model that may be applicable to other species. Laboratory experiments were used to determine per capita grazing rate (PCGR; g seaweeds/individual/day). Relationship between the biomass of individual grazers and fourth-root transformed PCGR was fitted to power curve ($y=0.2310x^{0.3290}$, r=0.8864). This finding supported that the grazing efficiency was not even as individual grazers increase in size (biomass). Therefore, the biomass-normalized PCGR was estimated and revealed that smaller size herbivores were more effective grazers. Grazing impact considering density of each taxon was calculated. The sea hare Aplysia kurodai had greatest grazing impact on the seaweed bed and the sea urchin Strongylocentrotus nudus and S. intermedius were ranked in descending order of the impact. The amount of seaweed grazed by the amphipod Elasmopus sp. (>4,000 $ind./m^2$) and Jassa falcata (>2,000 $ind./m^2$) were 3.435 and $1.697mg/m^2/day$ respectively. The combined grazing amount of herbivores was $5,045mg/m^2/day$ in the seaweed bed. Although sea hare and sea urchin had strong impacts on seaweeds, the effects of dense, smaller species could not be seen as negligible. Surprisingly, the calculated grazing potential of sea urchins with a mean density of 3 $ind./m^2$ exceeded the mean production of seaweed cultured in domestic coastal waters in Korea (ca., 5 ton/ha). Small crustaceans were also expected to consume up to 16% of the seaweed production if their densities were rising under weak predation conditions. Considering that the population density of herbivores are strongly controlled by fish, human interference like overfishing may have strong negative effects on persistence of seaweeds communities.

Phenylarsine Oxide and Adenosine-sensitive Trans-golgi Complex Targeting of GFP Fused to Modified Sulfatide-binding Peptide (Phenylarsine oxide와 adenosine에 민감한 sulfatide 결합 펩타이드의 trans-golgi network 타기팅)

  • Jun, Yong-Woo;Lee, Jin-A;Jang, Deok-Jin
    • Journal of Life Science
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    • v.28 no.2
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    • pp.162-169
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    • 2018
  • Many cytoplasmic proteins are targeted to the cytoplasmic membrane of the trans-Golgi network (TGN) via an N-terminal short helix. We previously showed that the 20 N-terminal amino acids of Aplysia phosphodiesterase 4 (ApPDE4) long form are sufficient for its targeting to the plasma membrane and the TGN. The N-terminus of the ApPDE4 long form binds to PI4P and sulfatide in vitro. Therefore, in order to decipher the roles of sulfatide in Golgi complex targeting, we examined the cellular localization of sulfatide-binding peptides. In this study, we found that enhanced green fluorescent protein (EGFP) fused to the C-terminus of modified sulfatide- and heparin-binding peptides (mHSBP-EGFP) was localized to the TGN. On the other hand, its mutant, in which tryptophan was replaced with an alanine, leading to the impairment of heparin and sulfatide binding, was localized to cytosol. We also found that the TGN targeting of mHSBP-EGFP is impaired by the treatment of antimycin A, phenylarsine oxide (PAO), and adenosine but not a high concentration of wortmannin. These results suggest that PAO and adenosine-sensitive kinases, including phosphatidylinositol 4-kinase II, may play key roles in the recruitment of mHSBP-EGFP.

Proteolytic Activity of the Crude Enzyme Extracted from the Digestive Tract of Marine Gastropods (해산복족류의 소화관조직중에 분포하는 단백질분해효소의 활성)

  • CHO Deuk Moon;PYEUN Jae Hyeung;BYUN Dae Seok;KIM Chang Yang
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.16 no.3
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    • pp.216-224
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    • 1983
  • This work was undertaken to obtain some characteristics of proteolytic enzyme of marine gastropods such as sea hare, Aplysia kurodai, top shell, Turbo cornutus, and abalone, Haliotis discus hannai. An influence of pH, temperature and some chemicals on proteolytic activity of the crude enzyme extracted from digestive tract of the samples was taken into account and the stability of the enzyme during the storage at low temperature was also discussed. In comparison of the activities of the crude enzyme from the samples to the optimum conditions, it was characterized that abalone has twice or the more times higher activities than the other two species of the gastropoda in the acid and weak acid region, while, in the alkaline region, sea hare has six or the more times higher acitivities than the other two species. The proteolytic activity was facilitated by $Mn^{2+}$, some reducing agents, EDTA and DTT, and inhibited by $Hg^{2+}$ and SDS, but the other chemicals were not significantly affected to the activity. The low temperature storage of the enzymes of sea hare and top shell at $0^{\circ}C\;or\;-20^{\circ}C$ was not affected to the enzymic activity under the optimum pH condition except in the alkaline region. On the other hand, the low temperature storage was brought about no significant effect on the activity of the enzymes extracted from abalone under the optimum condition of the weak acid region, but apparently influenced to the activity under the optimum condition of the acid and alkaline region.

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Protein and Amino Acid Compositions in Echiurid and Sea Hare Muscles (개불과 군소육의 단백질 및 아미노산 조성)

  • CHOI Yeung-Joon;HAN Young-Sil
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.18 no.6
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    • pp.550-556
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    • 1985
  • In order to evaluate the marine mollusc muscle as foodstuff not only from the biochemical aspect but also from the view point of food science, we have analyzed the protein and amino acid compositions of the echiurid (Urechis unicinctus) and sea hare (Aplysia kurodai) muscle. The protein quality of the muscles was also investigated using in vitro methods based on in vitro digestibility, predicted digestibility, computed PER (C-PER) and discriminant computed PER(DC-PER). The remarkable feature of the protein compositions of the both muscles was that water soluble protein occupied a large amount of the muscle protein with fairly lower contents of the salt soluble protein. From the analysis of SDS-PAG electrophoresis, the sarcoplasmic proteins in the echiurid and the sea hare muscles were composed of 15 and 10 subunits, respectively. The free amino acid compositions of the total amino acids in the echiurid and sea hare muscle were characterized with $75\%$ of glycine and alanine, and with $78\%$ of taurine, respectively. The amino acid anaylsis of both muscle proteins showed that the echiurid muscle was rich in glycine, aspartic acid, glutamic acid, arginine and lysine, but was poor in cysteine, while the sea hare muscle was rich in glycine, glutamic acid, aspartic acid and arginine, but was negligible in cysteine and tryptophan. In the total amino acid profiles of the freeze dried muscles in echiurid and sea hare, there was not found a significant difference compared to the amino acid compositions of the muscle proteins. Predicting the protein quality of the echiurid and sea hare muscles using the in vitro method, it was apparently low compared to the muscle protein of fishes.

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