• Title/Summary/Keyword: protein secretion

Search Result 813, Processing Time 0.03 seconds

Effect of Agarase Signal Peptide from Agarivorans albus YKW-34 on Protein Secretion in Escherichia coli (대장균에서 단백질 분비에 대한 Agarivorans albus YKW-34의 Agarase 시그널펩티드의 효과)

  • Lee, Joo-Young;Song, Dae-Geun;Son, Jin-Ki;Pan, Cheol-Ho
    • Journal of Applied Biological Chemistry
    • /
    • v.53 no.2
    • /
    • pp.105-107
    • /
    • 2010
  • To overcome the limitation of E. coli expression system such as inclusion body formation and disulfide bond failure, we tried to express the heterologous protein as a secreted form. We adopted agarase signal peptide (ASP; 23 amino acid residues) from Agarivorans albus YKW-34 which is one of marine bacteia. When we used ASP to express $\beta$-agarase, about 42% activity was detected in media.

Interaction Proteome Analysis of Xanthomonas Hrp Proteins

  • Jang, Mi;Park, Byoung-Chul;Lee, Do-Hee;Bae, Kwang-Hee;Cho, Sa-Yeon;Park, Hyun-Seok;Lee, Baek-Rak;Park, Sung-Goo
    • Journal of Microbiology and Biotechnology
    • /
    • v.17 no.2
    • /
    • pp.359-363
    • /
    • 2007
  • Because of the importance of the type III protein-secretion system in bacteria-plant interaction, its function in bacterial pathogenesis of plants has been intensively studied. To identity bacterial proteins interacting with Xanthomonas hrp gene products that are involved in pathogenicity, we performed the glutathione-bead binding analysis of Xanthomonas lysates containing GST-tagged Hrp proteins. Analysis of glutathione-bead bound proteins by 1-DE and MALDI-TOF has demonstrated that Avr proteins, RecA, and several components of the type III secretion system interact with HrpB protein. This proteomic approach could provide a powerful tool in finding interaction partners of Hrp proteins whose roles in host-pathogen interaction need further studies.

EFFECTS OF CIMATEROL (CL 263,780) ON GROWTH PERFORMANCE AND CARCASS QUALITY OF BROILERS FED ON DIFFERENT LEVELS OF DIETARY PROTEIN AND ENERGY

  • Kim, Y.Y.;Han, I.K.;Ha, J.K.;Choi, Y.J.;Lee, M.
    • Asian-Australasian Journal of Animal Sciences
    • /
    • v.4 no.4
    • /
    • pp.383-393
    • /
    • 1991
  • The present study was carried out to investigate the effect of cimaterol on growth performance, carcass quality and cellular functional activity of broilers as affected by the various protein and energy levels. In starter period (0-21 days) all chicks were fed the basal diet which contained approximately 23 % crude protein and 3200 kcal of metabolizable energy per kg of diet. The cimaterol was added during 22-49 days and during the period of 8th week the cimaterol was withdrawn. In finisher period (22-49 days), a $2{\times}2{\times}3$ factorial arrangement consisting of 2 levels of cimaterol (0 mg/kg, 0.25 mg/kg), 2 levels of protein (19%, 17%) and 3 levels of energy (3200, 2900, 2600 kcal/kg) was used. In the finisher period, the body weight gain and feed efficiency was improved by the supplementation of cimaterol. The high protein and high energy level with supplementation of cimaterol had showed the highest body weight gain and feed efficiency, without significant difference. The administration of cimaterol had no effects on percentage of abdominal fat content, giblet and neck. Eventhough the difference was not significant (p>0.05), carcass yield was improved slightly by the administration of cimaterol. The effect of cimaterol on carcass composition was clearly demonstrated that protein content of broilers was not increased (p>0.05) but fat content decreased significantly (p<0.05). The ultilization of nutrients in experimental diets was not significantly affected by feeding cimaterol compared to control group. The results of in vitro studies with liver and adipose tissue showed that cimaterol increased the lipolytic activities at 19% protein level whereas at 17% protein level this effect was variable. Lipogenic activities in liver and adipose tissue were not affected with the administration of cimaterol but the activities increased as energy decreased, particularly in liver tissue. In cell studies with acinar culture of liver tissues, cimaterol had no effect on protein synthetic activity but the parameter was increased at higher level of dietary protein and energy. Protein secretion in liver was increased by the supplementation of cimaterol. In addition, at high protein level the protein secretion was increased and has shown the highest values at medium energy level.

Effects of Ginsenosides on Acid Secretion in Gastric Cells Isolated from Human and Rabbit Gastric Mucosa (인체 및 토끼 위선세포에서 인삼사포닌의 위산분비 매개 신호전달체계에 미치는 영향)

  • Kim, Hye-Yeong;Kim, Sin-Il;Kim, Gyeong-Hwan
    • Journal of Ginseng Research
    • /
    • v.22 no.1
    • /
    • pp.22-31
    • /
    • 1998
  • Antiulcer effects of ginseng saponin, acidic polysaccharide and methanol extract of Panax ginseng in the patients and experimental animals were reported. Postulated action mechanisms of ginseng were histamine-Ht receptor blocking and increasing gastric blood flow In the present study, the effect of ginsenosides, the biologically active glycosides of ginseng, on gastric acid secretion was examined using gastric cells isolated from human and rabbit gastric mucosa. Ginseng saponin, ginsenoside $Rb_1$, $Rb_2$, $Rg_1$ and $Rh_2$ were tested in unstimulated as well as stimulated gastric cells. Histamine ($10^4$M) and 3-isobutyl-1-methylxanthine ($10^4$M) were used as secretagogues. To investigate the mechanism of ginsenosides on acid secretion, the levels of cAMP and cGMP were monitored in gastric cells. As a result, high concerltration(1mg/ml) of ginseng saponin showed 73-75% of stimulated acid secretion in control gastric cells. However, ginseng saponin had no effect on unstimulated acid secretion and the levels of cGMP and cAMP in gastric cells. Ginsenoside $Rb_1$, $Rb_2$ and $Rh_2$ significantly inhibited stimulated acid secretion. Gastric cGMP levels were increased by all ginsenosides tested while cAMP levels were increased by all ginsenosides in unstimulated state of gastric cells, but increased by ginsenosides ginsenoside $Rg_1$ and $Rh_2$in stimulated state of gastric cells. The results suggest that inhibition of ginseng saponin on gastric acid secretion represents a complex effect of individual ginsenosides, which produce a range of effect on acid secretion. The inhibition site of ginseng saponin on stimulated acid secretion is postulated as post cAMP levels in acid secretary pathway such as protein phosphorylation or proton pump. Nitric oxide may not be involved in the inhibitory effect of ginseng saponin on stimulated acid secretion.

  • PDF

Glucose regulated protein 78 promotes cell invasion via regulation of uPA production and secretion in colon cancer cells

  • Li, Zongwei;Zhang, Lichao;Li, Hanqing;Shan, Shuhua;Li, Zhuoyu
    • BMB Reports
    • /
    • v.47 no.8
    • /
    • pp.445-450
    • /
    • 2014
  • Glucose regulated protein 78 (GRP78) is frequently highly expressed in tumor cells, contributing to the acquisition of several phenotypic cancer hallmarks. GRP78 expression is also positively correlated with tumor metastasis, and promotes hepatocellular carcinoma cell invasion via increasing cell motility, however, other mechanisms involving the prometastatic roles of GRP78 remain to be elucidated. Here we report that forced GRP78 expression promotes colon cancer cell migration and invasion through upregulating MMP-2, MMP-9 and especially uPA production. These effects of GRP78 are mediated by enhancing the activation of ${\beta}$-catenin signaling. Interestingly, we identify that GRP78 interacts with uPA both in the cells and in the culture medium, suggesting that GRP78 protein is likely to directly facilitate uPA secretion via protein-protein interaction. Taken together, our findings demonstrate for the first time that besides stimulation of cell motility, GRP78 can act by increasing proteases production to promote tumor cell invasion.

Baicalin suppresses lipopolysaccharide-induced matrix metalloproteinase expression: action via the mitogen-activated protein kinase and nuclear factor κB-related protein signaling pathway

  • Ko, Seon-Yle
    • International Journal of Oral Biology
    • /
    • v.46 no.1
    • /
    • pp.51-59
    • /
    • 2021
  • Periodontal disease is an inflammatory disease that affects the destruction of the bone supporting the tooth and connective tissues surrounding it. Periodontal ligament fibroblasts (PDLFs) induce overexpression of matrix metalloproteinase (MMP) involved in periodontal disease's inflammatory destruction. Osteoclasts take part in physiological bone remodeling, but they are also involved in bone destruction in many kinds of bone diseases, including osteoporosis and periodontal disease. This study examined the effect of baicalin on proteolytic enzymes' production and secretion of inflammatory cytokines in PDLFs and RAW 264.7 cells under the lipopolysaccharide (LPS)-induced inflammatory conditions. Baicalin inhibited the expression of the protein, MMP-1 and MMP-2, without affecting PDLFs' cell viability, suggesting its possibility because of the inhibition of phosphorylation activation of mitogen-activated protein kinase's p38, and the signal transduction process of nuclear factor κB (NFκB)-related protein. Also, baicalin reduced the expression of MMP-8 and MMP-9 in RAW 264.7 cells. This reduction is thought to be due to the inhibition of the signal transduction process of NFκB-related proteins affected by inhibiting p65RelA phosphorylation. Also, baicalin inhibited the secretion of nitric oxide and interleukin-6 induced by LPS in RAW 264.7 cells. These results suggest that baicalin inhibits connective tissue destruction in periodontal disease. The inhibition of periodontal tissue destruction may be a therapeutic strategy for treating inflammatory periodontal-diseased patients.

Effect of Individual Fatty Acids on Synthesis and Secretion of Apolipoprotein and Lipoprotein in hep-G2 Cells

  • Ryowon Choue
    • Journal of Nutrition and Health
    • /
    • v.27 no.9
    • /
    • pp.910-923
    • /
    • 1994
  • The effects of individual fatty acids, differing in their degree of unsaturation(18:0, 18:1, 18:2 and 18:3) on the biosynthesis and secretion and lipids were investigated in Hep-G2 cells. Synthesis of apolipoprotein was measured by the incorporation of 3H-leucine into apolipoprotein(d<1.21g/ml) and synthesis of lipids was measured by the incorporation of 3H-glycerol and 14C-acetate into various lipid classes. Inclusion of 1.0mM of each fatty acids into the culture medium significantly increased the synthesis of total apolipoprotein and Apo B(p<0.05). However, addition of fatty acid did not affect the synthesis of cellular and medium protein. Among different fatty acids tested, oleic acid had the greatest effect on Apo B synthesis. While stearic, linoleic and linolenic acid, all had similar effects. The secretion of triglyceride into the medium markedly increased in all fatty acid groups being 5-6 times over the albumin control. The triglyceride secretion was the highest int he oleic acid group. The secretion of phospholipid and cholesterol also increased with triglyceride output. A positive relationship existed between the output of lipoprotein-triglyceride and Apo B. Since the synthesis of Apo B was significantly increased when various fatty acids were included into the culture medium, part of the apparently stimulated synthesis of the apolipoprotein may be in response to the increased formation and secretion of lipoprotein lipids.

  • PDF

The Glucoamylase Signal Sequence Directs the Efficient Secretion of Human $\alpha$1-Antitrypsin in Yeast Cells (효모에서 Glucoamylase 신호서열에 의한 인체 $\alpha$1-Antitrypsin의 분비 효율 향상)

  • Song, Moo-Young;Kwon, Ki-Sun;Kang, Dae-Ook;Yu, Myeong-Hee;Park, Hee-Moon;Kim, Jinmi
    • Korean Journal of Microbiology
    • /
    • v.31 no.3
    • /
    • pp.203-207
    • /
    • 1993
  • Five different secretion vectors were constructed by varying the signal sequences and .alpha.-antitrypsin (.alpha.1-AT) a numan secretory protein, was produced from yeast cells. The signal sequences used are those of acid phosphatase (PH05) and .alpha.-factor (M f.alphal1) of Saccharomyces cerevisiae, glucoamylase (STA1) of Saccharomyces diastaticus, and human .alpha.1-AT. Four vectors directed the efficient secretion of .alpha.1-AT ito the culture media. The secretion vector carrying the glucoamylase signal sequence (pGAT11) showed the highest efficiency of secretion. About 70% of .alpha.1-AT produce dwere secreted into the media. The endo H treatment of partially purified .alpha.1-AT indicates that the secreted .alpha.1-AT appeared to be glycosylated. This glycosylation pattern was altered when amino acid substitution mutations were introduced at the three glycosylation sites of .alpha.-AT.

  • PDF

Effect of oleanolic acid on the activity, secretion and gene expression of matrix metalloproteinase-3 in articular chondrocytes in vitro and the production of matrix metalloproteinase-3 in vivo

  • Kang, Dong-Geun;Lee, Hyun Jae;Kim, Kun Tae;Hwang, Sun-Chul;Lee, Choong Jae;Park, Jin Sung
    • The Korean Journal of Physiology and Pharmacology
    • /
    • v.21 no.2
    • /
    • pp.197-204
    • /
    • 2017
  • In the present study, we tried to examine whether oleanolic acid regulates the activity, secretion and gene expression of matrix metalloproteinase-3 (MMP-3) in primary cultured rabbit articular chondrocytes, as well as the production of MMP-3 in the knee joint of rat to evaluate the potential chondroprotective effect of oleanolic acid. Rabbit articular chondrocytes were cultured in a monolayer, and reverse transcription-polymerase chain reaction (RT-PCR) was used to measure interleukin-$1{\beta}$ (IL-$1{\beta}$)-induced gene expression of MMP-3, MMP-1, MMP-13, a disintegrin and metalloproteinase with thrombospondin motifs-4 (ADAMTS-4), ADAMTS-5 and type II collagen. In rabbit articular chondrocytes, the effects of oleanolic acid on IL-$1{\beta}$-induced secretion and proteolytic activity of MMP-3 were investigated using western blot analysis and casein zymography, respectively. The effect of oleanolic acid on in vivo MMP-3 protein production was also examined, after intra-articular injection to the knee joint of rat. The results were as follows: (1) oleanolic acid inhibited the gene expression of MMP-3, MMP-1, MMP-13, ADAMTS-4, and ADAMTS-5, but increased the gene expression of type II collagen; (2) oleanolic acid reduced the secretion and proteolytic activity of MMP-3; (3) oleanolic acid suppressed the production of MMP-3 protein in vivo. These results suggest that oleanolic acid can regulate the activity, secretion and gene expression of MMP-3, by directly acting on articular chondrocytes.