• Title/Summary/Keyword: G Proteins

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A MOLECULAR BIOLOGICAL STUDY ON THE EXPRESSION PATTERN AND FUNCTIONAL PROTEIN STRUCTURES OF PROLINE-RICH PROTEINS IN HUMAN SALIVARY GLANDS (사람의 타액선에서 proline-rich protein의 발현양상과 기능적 단백 구조에 대한 분자생물학적 연구)

  • Joo, Jae-Yong;Lee, Suk-Keun;Park, Young-Wook
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • v.28 no.1
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    • pp.31-41
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    • 2002
  • Proline-rich proteins (PRPs) are major components of human saliva. In order to know the biological roles of PRPs, we explored the expression pattern and functional protein structures of PRPs by the immunohistochemical and various molecular biological methods. Polyclonal antibody against human gPRP was generated from rabbit by the injection of oral exfoliated cells specially treated by urea and SDS buffer. The PRPs began to be expressed both in the acinar cells and ductal cells from the EIDS (Early Intermediate Developmental Stage) of fetal salivary glands and became intense in the salivary epithelium in the LDS (Late Developmental Stage) and adult salivary glands. The polyclonal antibody against the gPRP showed the cross-reactivity with aPRP and bPRP, these results were relevant to the high homology among subtypes of PRP. However, the simulated protein structures of PRPs showed the characteristic repetitive whorling domains except the N-terminal signal peptide. The whorling domains were also contained the multiple amino acids of glutamine and glycine, which may provide the receptor binding or cross-linking sites of PRPs.

Understanding of Protein Adsorption Kinetics to Contact Lens Hydrogels (콘택트렌즈용 하이드로젤로의 단백질 흡착 반응속도 이해)

  • Kim, Hyun-Jae;Kim, Mira;Noh, Hyeran
    • Polymer(Korea)
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    • v.38 no.2
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    • pp.220-224
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    • 2014
  • Protein adsorption kinetics was studied with the amount of proteins adsorbed to contact lens hydrogels over time scales. Hydroxyethylmethacrylate (HEMA) and silicone hydrogels were dipped in protein solutions (albumin or IgG) and adsorption amounts were measured over time scales. The amount of protein adsorbed to both hydrogel types increased rapidly in 10 min, and remained consistently in 90 min. Decreasing interfacial energetics was taken slowly up to an hour in spite of rapid diffusion of protein molecules. This is due to the fact that water deprivation from three dimensional interphase initially formed by protein diffusion took over an hour. Interpretation of adsorption kinetics on contact lens hydrogels was discussed with understanding of relationship between surface energy and protein adsorption capacity.

Protective Effect of Dodamtanggami-bang on Endoplasmic Reticulum Stress in C6 Glial Cells (도담탕가미방(導痰湯加味方)이 tunicamycin에 의한 소포체 스트레스성 C6 glial 세포사멸에 미치는 영향)

  • Kim, Bong-Sang;Moon, Byung-Soon
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.24 no.6
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    • pp.1004-1011
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    • 2010
  • This study was designed to evaluate the protective effects of Dodamtanggami-bang (DDTG) on tunicamycin induced cell death by ER stress in C6 glial cells. Cell viability was measured by MTT assay and LDH release. Apoptosis was determined by caspase activity and flow cytometry in C6 glial cells. Expression of ER stress mediators including, GRP78 and CHOP proteins were measured by Western blot analysis. Tunicamycin induced the apoptosis of C6 glial cells, which was characterized as nucleic acid and caspase-3 activation, PARP cleavage, and sub-G0/G1 fraction of cell cycle increase. However, pretreatment with DDTG protected C6 glial cells from tunicamycin. Treatment with tunicamycin resulted in the increased the expression of GRP78 and CHOP protein and produced ROS generation. However, pretreatment with DDTG inhibited the ER stress pathway, including increase of the expression of GRP78, CHOP proteins in C6 glial cells treated with tunicamycin. Taken together, these data suggest that DDTG is able to protect C6 glial cells from tunicamycin with marked inhibition of ER stress.

Cellular Effects of Troglitazone on YD15 Tongue Carcinoma Cells

  • Loan, Ta Thi;Yoo, Hoon
    • International Journal of Oral Biology
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    • v.41 no.3
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    • pp.113-118
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    • 2016
  • An FDA approved drug for the treatment of type II diabetes, Troglitazone (TRO), a peroxisome proliferator-activated receptor gamma agonist, is withdrawn due to severe idiosyncratic hepatotoxicity. In the search for new applications of TRO, we investigated the cellular effects of TRO on YD15 tongue carcinoma cells. TRO suppressed the growth of YD15 cells in the MTT assay. The inhibition of cell growth was accompanied by the induction of cell cycle arrest at $G_0/G_1$ and apoptosis, which are confirmed by flow cytometry and western blotting. TRO also suppressed the expression of cell cycle proteins such as cyclin D1, cdk2, cdk4, cyclin B1, cdk1(or cdc2), cyclin E1 and cyclin A. The inhibition of cell cycle proteins was coincident with the up-regulation of $p21^{CIP1/WAF1}$ and $p27^{KIP1}$. In addition, TRO induces the activation of caspase-3 and caspase-7, as well as the cleavage of PARP. Further, TRO suppressed the expressions of Bcl-2 without affecting the expressions of Bad and Bax. Overall, our data supports that TRO induces cell cycle arrest and apoptosis on YD15 cells.

Studies on the Chemical Composition of Korean Wheat Flour Proteins (I). Purification of Gliadin Proteins and Amino Acid Composition (한국산 밀가루 단백질의 화학적 조성에 관한 연구 (제1보). Gliadin 단백질의 아미노산 조성 및 그의 정제)

  • Sung Hye Cho;Kim Jun Pyong
    • Journal of the Korean Chemical Society
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    • v.21 no.3
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    • pp.210-214
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    • 1977
  • We have investigated the amino acid composition of gliadin and glutenin from wheat flour Jang Kwang variety and have purified the main gliadin protein by Sephadex column. The results were obtained as follows: 1. Five bands of component for both gliadin and glutenin were found in disc electrophoretic gel column 2. The highest content of amino acid in gliadin and glutenin were glutamic acid and alanine was present in comparatively large amounts in glutenin, whereas proline was rich in gliadin 3. Main gliadin protein was purified by Sephadex G-150 and A-25 column chromatography and identified its purity by disc electrophoresis.

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Statistical optimization of culture media contained soy proteins and hypocotyl for the growth of Bifidobacterium lactis BL 740 and production of soy isoflavone aglycones (대두 단백질 및 배아를 이용한 Bifidobacterium lactis BL740의 균체성장 및 이소플라본 비배당체 생산를 위한 통계적 배지 최적화)

  • Lee, Choong-Young;Lee, Yoon-Bok;Lee, Keun-Ha;Park, Myeong-Soo;Hwang, Seock-Yeon;Hong, Seung-Bok;Yoo, Yung-Choon;Yu, Byung-Yeon;Kim, Chung-Ho
    • Journal of Applied Biological Chemistry
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    • v.53 no.3
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    • pp.126-131
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    • 2010
  • In order to maximize the growth of Bifidobacterium lactis BL 740 and soy isoflavone agycones production, we investigated the optimization of a culture medium containing soy hypocotyls, which are the byproducts of the soy manufacturing process, and soy proteins. The ingredients of the medium containing soy materials (S-medium) were selected by fractional factorial design (FFD) and central composite design (CCD) within a desirable range. The FFD was applied by six factors: glucose, cellobiose, fructooligosaccharide, soy peptone, soy protein, and soy hypocotyl. Soy protein, soy peptone, and soy hypocotyl were found to be significant factors from the result of FFD for both the growth of B. lactis BL 740 and aglycone production. The CCD was then applied with three variables found from FFD at five levels each and the optimum values were determined for the three variables: soy peptone, soy protein, and soy hypocotyl. In the case of the growth of B. lactics BL740, the proposed optimal media contained 12.73 g/L of soy protein, 29.55 g/L of soy peptone, and 130.67 g/L of soy hypocotyl. To produce isoflavone aglycones, optimized media was composed of 2.06 g/L, soy protein, 1.25 g/L of soy peptone, and 60.02 g/L of soy hypocotyl.

Keratin 17 identified by proteomic analysis may be involved in tumor angiogenesis

  • Xu, Yong;Zhang, Su-Zhen;Huang, Can-Hua;Liu, Xin-Yu;Zhong, Zhen-Hua;Hou, Wen-Li;Su, Zi-Fen;Wei, Yu-Quan
    • BMB Reports
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    • v.42 no.6
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    • pp.344-349
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    • 2009
  • Angiogenesis is crucial for solid tumor growth. By secreting angiogenic factors, tumor cells induce angiogenesis. However, targeting these angiogenic factors for cancer therapy is not always successful, suggesting that other factors may be involved in tumor angiogenesis. This work shows that 25 protein spots were differentially expressed by two-dimensional gel electrophoretic analysis when HepG2 cells induced endothelial cell differentiation to tube in vitro, and most of them were upregulated. Twenty-one proteins were identified with MALDITOF-MS, and the other four were identified by LTQ-MS/MS. Keratins were identified as one class of these upregulated proteins. Further study indicated that the expression of keratin 17 in cultured endothelial cells is likely microenvironment regulated, because its expression can be induced by HepG2 cells and bFGF as well as serum in culture media. Increased expression of keratins in endothelial cells, such as keratin 17, may contribute to the angiogenesis induced by HepG2 cells.

Blocking Bcl-2 Leads to Autophagy Activation and Cell Death of the HEPG2 Liver Cancer Cell Line

  • Du, Peng;Cao, Hua;Wu, Hao-Rong;Zhu, Bao-Song;Wang, Hao-Wei;Gu, Chun-Wei;Xing, Chun-Gen;Chen, Wei
    • Asian Pacific Journal of Cancer Prevention
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    • v.14 no.10
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    • pp.5849-5854
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    • 2013
  • Background: Apoptosis may be induced after Bcl-2 expression is inhibited in proliferative cancer cells. This study focused on the effect of autophagy activation by ABT737 on anti-tumor effects of epirubicin. Methods: Cytotoxic effects of ABT737 on the HepG2 liver cancer cell line were assessed by MTT assay and cell apoptosis through flow cytometry. Mitochondrial membrane potential was measured by fluorescence microscopy. Monodansylcadaverin (MDC) staining was used to detect activation of autophagy. Expression of p53, p62, LC3, and Beclin1, apoptotic or autophagy related proteins, was detected by Western blotting. Results: ABT737 and epirubicin induced growth inhibition in HepG2 cells in a dose- and time-dependent manner. Both ABT737 and epirubicin alone could induce cell apoptosis with a reduction in mitochondrial membrane potential as well as increased apoptotic protein expression. Further increase of apoptosis was detected when HepG2 cells were co-treated with ABT373 and epirubicin. Furthermore, our results demonstrated that ABT373 or epirubicin ccould activate cell autophagy with elevated autophagosome formation, increased expression of autophagy related proteins and LC3 fluorescent puncta. Conclusions: ABT737 influences cancer cells through both apoptotic and autophagic mechanisms, and ABT737 may enhance the effects of epirubicin on HepG2 cells by activating autophagy and inducing apoptosis.

Characterization and Condition of Silver Accumulation Bacteria in Groundwater (지하수에서 분리한 은(銀) 축적균주에 대한 축적조건 및 특성)

  • 배진희;민병례;한명수;최영길
    • Korean Journal of Microbiology
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    • v.37 no.4
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    • pp.273-276
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    • 2001
  • The strain which accumulate the silver in cell were isolated and characterized. And condition of accumulation of heavy metal was examined closely to investigate optimal condition of accumulation. Pseudomonas fluorescens and Bacillus cereus were Isolated from groundwater as the strain of silver accumulating bacteria. These strains did not grow in the medium at silver over the concentration 20 ppm. The largest accumulations of silver in the culture of Pseudomonas fluorescens and Bacillus cereus occurred within 24 hours. The amount of silver accumulation in Pseudomonas fluorescens and Bacillus cereus were 1.9 mg/g cell and 1.65 mg/g cell, repletively. In protein patterns of cell after the treatment of silver, three reducible proteins (126 KDa, 89 KDa, 25 KDa) in Bacillus cereus and one new protein (34 KDa) in Pseudomonas fluorescens were detected by SDS-PAGE.

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Differential Modulatory Effects of Cholera Toxin and Pertussis Toxin on Pain Behavior Induced by TNF-${\alpha}$, Interleukin-1${\beta}$ and Interferon-${\gamma}$ Injected Intrathecally

  • Kwon, Min-Soo;Shim, Eon-Jeong;Seo, Young-Jun;Choi, Seong-Soo;Lee, Jin-Young;Lee, Han-Kyu;Suh, Hong-Won
    • Archives of Pharmacal Research
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    • v.28 no.5
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    • pp.582-586
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    • 2005
  • The present study was designed to characterize the possible roles of spinally located cholera toxin (CTX)- and pertussis toxin (PTX)-sensitive G-proteins in pro- inflammatory cy tokine induced pain behaviors. Intrathecal injection of tumor necrosis factor-a (TNF-${\alpha}$; 100 pg), interleukin-1${\beta}$ (IL-1${\beta}$ 100 pg) and interferon-${\gamma}$ (INF-${\gamma}$; 100 pg) showed pain behavior. Intrathecal pretreatment with CTX (0.05, 0.1 and 0.5 mg) attenuated pain behavior induced by TNF-${\alpha}$ and INF-${\gamma}$ administered intrathecally. But intrathecal pretreatment with CTX (0.05, 0.1 and 0.5${\mu}g$) did not attenuate pain behavior induced by IL-1${\beta}$. On the other hand, intrathecal pretreatment with PTX further increased the pain behavior induced by TNF-${\alpha}$ and IL-1${\beta}$ administered intrathecally, especially at the dose of 0.5 ${\mu}g$. But intrathecal pretreatment with PTX did not affect pain behavior induced by INF-${\gamma}$. Our results suggest that, at the spinal cord level, CTX- and PTX-sensitive G-proteins appear to play important roles in modulating pain behavior induced by pro-inflammatory cytokines administered spinally. Furthermore, TNF-${\alpha}$, IL-1${\beta}$ arid INF-${\gamma}$ administered spinally appear to produce pain behavior by different mechanisms.