• Title/Summary/Keyword: G protein $G{\alpha}_{13}$

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Gene Expression Analysis of Gα13-/- Knockout Mouse Embryos Reveals Perturbations in Gα13 Signaling Related to Angiogenesis and Hypoxia

  • Park, Ji-Hwan;Choi, Sang-Dun
    • Genomics & Informatics
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    • v.9 no.4
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    • pp.161-172
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    • 2011
  • Angiogenesis is regulated by a large number of molecules and complex signaling mechanisms. The G protein $G{\alpha}_{13}$ is a part of this signaling mechanism as an endothelial cell movement regulator. Gene expression analysis of $G{\alpha}_{13}$ knockout mouse embryos was carried out to identify the role of $G{\alpha}_{13}$ in angiogenesis signaling during embryonic development. Hypoxia-inducible response factors including those acting as regulators of angiogenesis were over expressed, while genes related to the cell cycle, DNA replication, protein modification and cell-cell dissociation were under expressed. Functional annotation and network analysis indicate that $G{\alpha}_{13}{^{-/-}}$ embryonic mice were exposed to hypoxic conditions. The present analysis of the time course highlighted the significantly high levels of disorder in the development of the cardiovascular system. The data suggested that hypoxia-inducible factors including those associated with angiogenesis and abnormalities related to endothelial cell division contributed to the developmental failure of $G{\alpha}_{13}$ knockout mouse embryos.

The Changes of $\alpha$-galactosidase Activities and Stachyose and Raffinose Contents During Fermentation of Soybeans (대두의 발효에 따른 $\alpha$-Galactosidase활성 및 Stachyose, Raffinose 함량 변화)

  • Kim, Sung-Soo;Yoon, Sun
    • Korean journal of food and cookery science
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    • v.14 no.5
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    • pp.509-512
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    • 1998
  • Changes in the contents of stachyose and raffinose were determined during soybean fermentation. ${\alpha}$-Galactosidase activities were also monitored in soybean and its fermented products. The stachyose contents were 31.8239 mg/g of soybean, 4.2217 mg/g of Meju, and 2.1184 mg/g of Doenjang. The raffinose contents were 2.6914 mg/g of soybean, 1.7413 mg/g of Meju, and negligible of Doenjang. ${\alpha}$-Galactosidase activities was distinct in soybean and Meju. They were 14.5954 units/mg protein of soybean, 13.1489 units/mg protein of Meju, and 1.9157 units/mg protein of Doenjang. The results suggested that the decrease of stachyose and raffinose contents in fermented soy products were due to the ${\alpha}$-galactosidase activity.

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Potential drug targets in the GPCR-$G{\alpha}_{12}/G{\alpha}_{13}$ signaling pathways

  • Kim, Sang-Geon
    • Proceedings of the Korean Society of Applied Pharmacology
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    • 2008.04a
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    • pp.89-99
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    • 2008
  • GPCRs are large families of cell surface receptors that transmit signals through conformational changes upon ligand activation and an interaction with the heterotrimeric G-proteins. GPCRs regulate the cell-signaling pathways and participate in the regulation of physiological processes through the G-proteins defined by their ${\alpha}$ subunits. A family of 20 G protein-coupled receptors (GPCRs) that provide a large class of tractable drug targets for new anti-inflammatory drugs and, in certain instances, for the treatment of the inflammatory indications such as atherosclerosis, rhinitis, asthma, pulmonary disease and arthritis. In view of the important findings showing that $G{\alpha}_{12}/G{\alpha}_{13}$ regulate the various cellular processes such as actin-stress fiber formation, neurite retraction, platelet aggregation, gene induction, and apoptosis, we became interested in whether, after coupling to the activated GPCRs, the G-proteins and their downstream molecules might be involved in the pathologic processes of chronic inflammatory diseases (e.g., liver fibrosis). In this symposium, the possible link of the G-proteins with the pathophysiology will be discussed with the aim of finding potential new drug targets.

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Effect of Coptidis Rhizoma Extract on Cytokine Production of Mouse Macrophages (황연(黃連) 추출물이 대식세포의 면역단백질 생성에 미치는 영향)

  • Kim, Bok-Kee;Han, Hyo-Sang;Lee, Young-Jong
    • Herbal Formula Science
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    • v.21 no.2
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    • pp.81-89
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    • 2013
  • Objectives : The purpose of this study is to observe the effect of Coptidis Rhizoma (CCE-extract of C. chinensis Rhizome) in induction of immune protein on mouse macrophages. Methods : To analyze cytokines interleukin(IL)-$1{\alpha}$, IL-3, IL-9, IL-12p40, IL-13, IL-17, Monocyte Chemoattractant Protein(MCP)-1 induced by macrophages, mouse macrophages were incubated with CCE and was measured. Results : IL-$1{\alpha}$ measurement, CCE showed significant inhibition only at concentration level of 200 ${\mu}g/mL$. IL-3, MCP-1 measurement, CCE showed significant inhibition only at concentration level of 100, 200 ${\mu}g/mL$. IL-9 measurement, CCE showed significant inhibition only at concentration level of 50 ${\mu}g/mL$. IL-13 measurement, CCE showed significant inhibition only at concentration level of 50, 100, 200 ${\mu}g/mL$. The IL-12p40, IL-17 levels indicated no changes at 25, 50, 100, 200 ${\mu}g/mL$ on mouse macrophages. Conclusions : CCE did not significantly increased inflammatory cytokines IL-$1{\alpha}$, IL-3, IL-9, IL-12p40, IL-13, IL-17, Monocyte Chemoattractant Protein(MCP)-1 on mouse macrophages. It was verified CCE does not trigger cytokine related hypersensitivity reaction of organism or exacerbation of acute/chronic inflammatory disease.

The development of new soybean strain with ti and cgy1 recessive allele

  • Choi, Sang Woo;Park, Jun Hyun;Chung, Jong Il
    • Journal of Plant Biotechnology
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    • v.45 no.4
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    • pp.328-332
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    • 2018
  • Soybean [Glycine max (L.) Merr.] seed is an important dietary source of protein, oil, carbohydrate, isoflavone and other various nutrients for humans and animals. However, there are anti-nutritional factors in the raw mature soybeans. Kunitz trypsin inhibitor (KTI) protein and stachyose are the main anti-nutritional factors in soybean seed. The ${\alpha}^{\prime}$-subunit of ${\beta}$-conglycinin protein exhibit poor nutritional and food processing properties. The genetic removal of the KTI and ${\alpha}^{\prime}$-subunit proteins will improve the nutritional value of the soybean seed. The objective of this research was to develop a new soybean strain with KTI and ${\alpha}^{\prime}$-subunit protein free ($titicgy_1cgy_1$ genotype) and proper agronomic traits. A breeding population was developed from the cross of the Bl-1 and 15G1 parents. A total of 168 $F_2$ seeds from the cross of the BL-1 and 15G1 parents were obtained. The segregation ratios of 9: 3: 3: 1 ($104Ti\_Cgy_{1\_}:\;30Ti\_cgy_1cgy_1:\;21cgy_1cgy_1Ti\_:\;13titicgy_1cgy_1$) between the Ti and $Cgy_1$ genes in the $F_2$ seeds were observed (${\chi}^2=5.12$, P=0.5-0.10). Two $F_4$ plant strains with proper agronomical traits and $titicgy_1cgy_1$ genotype (free of both KTI and ${\alpha}^{\prime}$-subunit protein) were selected and harvested. 2 strains (S1 and S2) had yellow seed coats and hilum. The plant height of the S1 strain was 65 centimeters. The 100-seed weight was 29.2 g. The plant height of the S2 strain was 66 centimeters and 100-seed weight was 26.2 g. The two strains selected in this research will be used to improve the new cultivar that will be free of the KTI and ${\alpha}^{\prime}$-subunit proteins.

Suppressive Effects of Defatted Green Tea Seed Ethanol Extract on Cancer Cell Proliferation in HepG2 Cells (HepG2 Cell에서 녹차씨박 에탄올 추출물의 암세포 증식 억제효과)

  • Noh, Kyung-Hee;Min, Kwan-Hee;Seo, Bo-Young;Kim, Hye-Ok;Kim, So-Hee;Song, Young-Sun
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.40 no.6
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    • pp.767-774
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    • 2011
  • Defatted green tea seed was extracted with 100% ethanol for 4 hr and then fractionated with petroleum ether, ethyl acetate and butanol. The ethanol and butanol extracts showed greater increases in antiproliferation potential against liver cancer cells than petroleum ether, ethyl acetate, $H_2O$, and hot water extracts did. Thus, this study was carried out to investigate the anti-proliferative actions of defatted green tea seed ethanol extract (DGTSE) in HepG2 cancer cells. The DGTSE contained catechins including EGC ($1039.1{\pm}15.2\;g/g$), tannic acid ($683.5{\pm}17.61\;{\mu}g/g$), EC ($62.4{\pm}5.00\;{\mu}g/g$), ECG ($24.4{\pm}7.81\;{\mu}g/g$), EGCG ($20.9{\pm}0.96\;{\mu}g/g$) and gallic acid ($2.4{\pm}0.68\;{\mu}g/g$), but caffeic acid was not detected when analyzed by HPLC. The anti-proliferation effect of DGTSE toward HepG2 cells was 83.13% when treated at $10\;{\mu}g$/mL, of DGTSE, offering an $IC_{50}$ of $6.58\;{\mu}g$/mL. DGTSE decreased CYP1A1 and CYP1A2 protein expressions in a dose-dependent manner. Quinone reductase and antioxidant response element (ARE)-luciferase activities were increased about 2.6 and 1.94-fold at a concentration of $20\;{\mu}g$/mL compared to a control group, respectively. Enhancement of phase II enzyme activity by DGTSE was shown to be mediated via interaction with ARE sequences in genes encoding the phase enzymes. DGTSE significantly (p<0.05) suppressed prostaglandin $E_2$ level, tumor necrosis factor-${\alpha}$ (TNF-${\alpha}$) protein expressions, and NF${\kappa}$B translocation, but did not affected nitric oxide production. From the above results, it is concluded that DGTSE may ameliorate tumor and inflammatory reactions through the elevation of phase II enzyme activities and suppression of NF${\kappa}$B translocation and TNF-${\alpha}$ protein expressions, which support the cancer cell anti-proliferative effects of DGTSE in HepG2 cells.

Busulfan-Induced IgG-Protein Complex of Germ Cells and Its Utility for Selection of Spermatogonial Stem Cells

  • 주학진;천영신;권득남;김진회
    • Proceedings of the KSAR Conference
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    • 2001.03a
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    • pp.38-38
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    • 2001
  • Spermatogonial stem cells은 sperrnatogenesis에서 중요한 역할을 하며, 곡세정관의 기저막에 위치하고 있는 것으로 알려져 있다. 그러나, 그 동안 이 세포에 특이하게 발현되는 marker가 거의 알려져 있지 않아 spermatogonial stem cell의 연구에 많은 어려움을 가져왔다. 최근 일반적인 stem cell이 갖는 특성 중, 기저막과 상호작용을 하는 surface protein으로 integrin이 존재한다는 사실을 이용하여, anti-$\alpha$$_{6}$/ 또는 anti-$\beta$$_1$ integrin항체로 germ cell을 선발하여 정소에 이식한 결과, 높은 효율로 이식세포유래의 정자발생이 가능하다는 결과가 보고되었다 (Shinohara et al., 1999). 한편, 항암제의 일종인 busulfan을 마우스에 투여(40mg/kg)한 후 4-5주가 경과하면 세정관의 기저막에 위치하는 spermatogonia를 제외하고 대부분의 생식세포는 소멸한다 본 실험의 목적은 이러한 사실들을 이용하여 spermatogonial stem cell의 특성을 밝히고, 이 생식세포를 보다 간편하고 손쉽게 선발할 수 있는 시스템을 확립하는데 있다. Busulfan처리 후 5주가 경과된 마우스와 정상적인 13주령의 마우스 testis로부터 세포를 분리한 후 FITC-conjugated anti-IgG를 이용한 면역형광법으로 측정.분석한 결과, 형광표식된 세포비율이 대조군과 비교하여 busulfan을 처리한 경우에서 유의적인 증가를 보였다.(17$\pm$3.8%. 0.7$\pm$0.3% busulfan vs control). 또한, IgG와 결합한 단백질이 존재하는 이들 세포들은 곡세정관의 기저막을 따라 위치하며, 단백질과 복합체를 형성한 IgG는 anti-Ig $G_{2a}$와 반응하지 않는다는 사실을 관찰했다. 이러한 IgG 복합체를 형성한 세포들의 특성을 이용하여, IgG와 반응을 하지 않는 것으로 확인된 이차 항체인 an1i-Ig $G_{2}$와 일차 항체인 anti-$\alpha$$_{6}$ 또는 anti-$\beta$$_1$ integrin항체를 이용하여 측정.분석하였다. Busulfan을 처리한 마우스 정소에서 분리한 세포를 다시 laminin으로 코팅된 dish에서 선발.회수해서, anti-lgG, anti-$\alpha$$_{6}$ 또는 anti-$\beta$$_1$ integrin항체로 각각 표식된 세포비율을 비교하였다. Laminin으로부터 선발.회수한 세포에서는 IgG복합체가 $\alpha$$_{6}$ 또 는 $\beta$$_1$integrin과 거의 같은 수준에서 높은 비율로 표식되었다. 결론적으로, busulfan에 의해 유도된 IgG와 결합가능한 단백질은 $\alpha$$_{6}$$\beta$$_1$ integrin과 마찬가지로 immunoglobulin G를 이용하여 spermatogonial stem cell의 선발을 가능하게 했다. 따라서, busulfan처리시 IgG는 미분화된 정조세포의 선발을 위한 하나의 marker로서 사용가능함을 시사한다.다.

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Anti-Inflammatory and Anti-Fibrotic Activities of Nocardiopsis sp. 13G027 in Lipopolysaccharides-Induced RAW 264.7 Macrophages and Transforming Growth Factor Beta-1-Stimulated Nasal Polyp-Derived Fibroblasts

  • Choi, Grace;Kim, Geum Jin;Choi, Hyukjae;Choi, Il-Whan;Lee, Dae-Sung
    • Microbiology and Biotechnology Letters
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    • v.49 no.4
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    • pp.543-551
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    • 2021
  • Nocardiopsis species produce bioactive compounds, such as antimicrobial and anti-cancer agents and toxins. However, no reports have described their anti-inflammatory and anti-fibrotic effects during nasal polyp (NP) formation. In this study, we investigated whether marine-derived bacterial Nocardiopsis sp. 13G027 exerts anti-inflammatory and anti-fibrotic effects on lipopolysaccharide (LPS)-stimulated RAW 264.7 macrophages and transforming growth factor (TGF)-β1-induced NP-derived fibroblasts (NPDFs). Nitric oxide (NO) and prostaglandin E2 (PGE2) levels were analyzed. Extract from Nocardiopsis sp. 13G027 significantly inhibited the upregulation of NO and PGE2 in LPS-activated RAW 264.7 macrophages. The expression of mitogen-activated protein kinases (MAPKs) and protein kinase B (Akt/PKB) in LPS-induced RAW 264.7 macrophages was evaluated; smooth muscle alpha-actin (α-SMA), collagen type I (Col-1), and fibronectin also phosphorylated small mothers against decapentaplegic (SMAD) 2 and 3 in TGF-β1-stimulated NPDFs. The Nocardiopsis sp. 13G027 extract suppressed the phosphorylation of MAPKs and Akt and the DNA-binding activity of activator protein 1 (AP-1). The expression of pro-fibrotic components such as α-SMA, Col-1, fibronectin, and SMAD2/3 was inhibited in TGF-β1-exposed NPDFs. These findings suggest that Nocardiopsis sp. 13G027 has the potential to treat inflammatory disorders, such as NP formation.

Ric-8B Modulates the Function of Alpha Subunit of Go

  • Kim, Seung-Hyun;Ghil, Sung-Ho
    • Biomedical Science Letters
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    • v.13 no.2
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    • pp.127-133
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    • 2007
  • Heterotrimeric GTP binding proteins (G proteins) mediate signal generated by neurotransmitter and hormones. Among all G proteins, Go is the most abundant in brain but its role in brain is not clearly understood. To determine the function of the alpha subunit of Go (Go$\alpha$), we search for the interacting partner of Go$\alpha$ in brain using yeast two-hybrid system. A resistant to inhibitor of cholinesterase (Ric-8B) was identified as a Go$\alpha$ interacting protein. We confirmed interaction between Go$\alpha$ and Ric-8b employing in vitro affinity binding assay and showed that the Ric-8b increased the function of Go$\alpha$. Our findings indicate that Ric-8b is possible guanine nucleotide exchange factor for Go$\alpha$.

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Proximate Composition, Cholesterol and $\alpha$-Tocopherol Content in 72 Species of Korean Fish (72종 어류의 일반성분조성과 콜레스테롤 및 $\alpha$-토코페롤함량)

  • JEONG Bo-Young;CHOI Byeong-Dae;LEE Jong-Soo
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.31 no.2
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    • pp.160-167
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    • 1998
  • Proximate composition, cholesterol (CHOL) and $\alpha-tocopherol$ ($\alpha-Toc$) content of seventy-two species of fish muscle (sea water fish, 59 species; fresh water fish, 13 species), which caught off Tongyeong coast of the Southern Sea ( Nam-Hae) and caught Kyeongho river, Sancheong, Korea, were determined. Lipid content was ranked from $0.53\%$ to $20.4\%$ and was high in the following order; mid-surface dwelling and migratory fish ($6.09{\pm}4.25\%$) > fresh water fish ($4.40{\pm}5.90\%$) > coastal and reep dwelling fish ($3.41{\pm}2.68\%$) > demersal fish ($2.12{\pm}2.41\%$). There was a negative correlation between the lipid and moisture content in all fish groups (p< 0.001). Protein was approximately $17\~18\%$ in all fish group, and no correlation was between the protein and moisture content in all but the demersal fish, which there was a negative correlation between both contents. Ash content was almost constant in all fish, accounting for $1\~2\%$, Cholesterol (CHOL) content was shown a great difference according to the fish species from 6.7 mg/100 g muscle (grass fish) to 249 mg/100 g (eel) and there was a positive correlation between the lipid and CHOL content. $\alpha-Toc$ content was less than 3 mg/100 g muscle in all fish, except that of eel (4.1 mg/100 g) and was relatively rich in the fish species contained a large amount of lipid.

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