• Title/Summary/Keyword: secretory protein

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Constitutive Exocytosis of Seminal Fluid Proteins in Male Accessory Glands and Ejaculatory Duct of Drosophila melanogaster

  • Jo, Gyeong-Sang;Kim, Seong-Yun;Im, Jeom-Hui;Jeong, Gi-Hwa;Lee, Cheong-Ju
    • Animal cells and systems
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    • v.1 no.3
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    • pp.515-520
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    • 1997
  • Male accessory glands and ejaculatory duct of Drosophila melanogaster are reproductive organs which synthesize secretory seminal proteins. Several products of these organs involved in egg laying, receptivity, and sperm stability or storage were isolated from their lumens. Despite their secretory process play an important role, exocytosis pathway in these organs is not well known. In the present study, we characterized secretory protein profiles and determined their secretory mechanisms. Eight accessory gland secretory proteins and two ejaculatory duct secretory proteins were detected in their lumens. All these proteins were constitutively synthesized in these organs and secreted to their lumens. Secretion of newly synthesized proteins initiated at about 1 h after synthesis, and reached the peak at 4 h after synthesis. It seems that secretion of the proteins may occur via constitutive exocytosis pathway.

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Cytokine Production in Cholangiocarcinoma Cells in Response to Clonorchis sinensis Excretory-Secretory Products and Their Putative Protein Components

  • Pak, Jhang Ho;Lee, Ji-Yun;Jeon, Bo Young;Dai, Fuhong;Yoo, Won Gi;Hong, Sung-Jong
    • Parasites, Hosts and Diseases
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    • v.57 no.4
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    • pp.379-387
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    • 2019
  • Clonorchis sinensis is a carcinogenic human liver fluke that promotes hepatic inflammatory environments via direct contact or through their excretory-secretory products (ESPs), subsequently leading to cholangitis, periductal fibrosis, liver cirrhosis, and even cholangiocarcinoma (CCA). This study was conducted to examine the host inflammatory responses to C. sinensis ESPs and their putative protein components selected from C. sinensis expressed sequenced tag (EST) pool databases, including $TGF-{\beta}$ receptor interacting protein 1(CsTRIP1), legumain (CsLeg), and growth factor binding protein 2 (CsGrb2). Treatment of CCA cells (HuCCT1) with the ESPs or bacterial recombinant C. sinensis proteins differentially promoted the secretion of proinflammatory cytokines ($IL-1{\beta}$, IL-6, and $TNF-{\alpha}$) as well as anti-inflammatory cytokines (IL-10, $TGF-{\beta}1$, and $TGF-{\beta}2$) in a time-dependent manner. In particular, recombinant C. sinensis protein treatment resulted in increase (at maximum) of ~7-fold in $TGF-{\beta}1$, ~30-fold in $TGF-{\beta}2$, and ~3-fold in $TNF-{\alpha}$ compared with the increase produced by ESPs, indicating that CsTrip1, CsLeg, and CsGrb2 function as strong inducers for secretion of these cytokines in host cells. These results suggest that C. sinensis ESPs contribute to the immunopathological response in host cells, leading to clonorchiasis-associated hepatobiliary abnormalities of greater severity.

Immunochemical Localization of Tetrahydrocannabinol (THC) in Chemically Fixed Glandular Thrichomes of Cannabis (Cannabaceae)

  • Eun Soo Kim;Paul G. Mahlberg
    • Animal cells and systems
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    • v.3 no.2
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    • pp.215-219
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    • 1999
  • Monoclonal antibody for delta-9-tetrahydrocannabiol (THC Ab), conjugated with protein A-gold, was employed as a probe to detect THC localization in the gland and subjacent cells of chemically fixed bracts of Cannabis. THC was detected in the outer wall of the disc cells, fibrillar matrix, the surface feature of secretory vesicles, and sheath throughout development of the secretory cavity. The probe was absent from vesicles. Label was also present in anticlinal walls of disc cells and walls of dermal and mesophyll cells. Little or no THC Ab was present in disc cells and none were detected in control tissues. This distribution pattern of THC Ab was similar to that in tissues prepared by high pressure cryofixation-cryosubstitution. Consistent association of THC with wall and wall-derived materials suggests that cannnabinoids are synthesized outside the plasma membrane and bound to a wall component, where-upon they are transported to the cavity with wall materials released from the disc cell wall during development of the secretory cavity.

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Bacillus subtilis의 단백질 분비기구 SecY의 유전자 수준의 조절이 단백질 분비에 미치는 영향

  • 김상숙;김순옥;서주원
    • Microbiology and Biotechnology Letters
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    • v.24 no.4
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    • pp.408-414
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    • 1996
  • The SecY is a central component of the protein export machinery that mediate the translocation of secretory proteins across the plasma membrane, and has been known to be rate-limiting factor of secretion in Escherichia coli. In order to study the extracellular protein secretion in Gram-positive microorganism, we have, constructed strains harboring more than one copy of the gene for SecY. Firstly, the gene, for B. subtilis SecY and its promoter region was subcloned into pDH32 and the chimeric vector was inserted into amyE locus by homologous recombination. Secondly, low copy number vector, pCED6, was also used for subcloning the secY gene and for constructing a strain which harbors several copies of secY. The KH1 cell which harbor two copies of secY on the chromosome excreted more extracellular proteins than the wild type PB2. Moreover, the KH2 cells which harbor several copies of secY in pCED6 vector excreted more extracellular proteins than the KH1 cells. Here, we found that the capacity of protein secretion is partly controlled by the number of secY and it is suggested that SecY has also an important role in protein secretion in B. subtilis, a gram positive microorganism, as like in E. coli. This will promote the use of B. subtilis as a host for the expression of useful foreign gene and excretion of precious proteins.

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Investigation of post-translational modification of the secreted protein expressed in insect cell lines using baculovirus expression vector system(BEVS)

  • Yun, Eun-Young;Goo, Tae-Won;Kim, Sung-Wan;Park, Kwang-Ho;Hwang, Jae-Sam;Kang, Seok-Woo;Kwon, O-Yu
    • Proceedings of the Korean Society of Sericultural Science Conference
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    • 2003.04a
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    • pp.82-83
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    • 2003
  • In previous experiment, we reported when the heterologous protein is expressed by using baculovirus expression vector system (BEVS), although the amount of intracellular protein is abundant, the amount of extracellular Protein is poor. As the link in the chain of the research, we investigated the secretory pathway, important in case of the secretory protein, of the protein expressed in insect cells using BEVS. (omitted)

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Molecular Characterization of a Bombyx mori Protein Disulfide Isomerase(bPDI) (누에 배양세포로부터 분리한 Protein Disulfide Isomerase 유전자의 발현 특성)

  • 구태원;윤은영;황재삼;강석우;권오유
    • Journal of Life Science
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    • v.11 no.5
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    • pp.415-422
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    • 2001
  • Many secreted proteins have disulfide bonds that are important for their structure and function. Protein disulfide isomerase (PDI, EC 5.3.1.4.), an enzyme that catalyzes the formation and rearrangement of thiol/disulfide exchange reactions, is a resident of the endoplasmic reticulum (ER). The subcellular localization and its function as catalyst of disulfide bond formation in the biosynthesis of secretory and cell membrane proteins suggest that PDI plays a key role in the secretory pathway. We have isolated a cDNA encoding protein disulfide isomerase from Bombyx mori(bPDI). It has been characterized under ER stress conditions (dominantly induced by calcium ionophore A23187, tunicamycin and DTT), which is known to cause an accumulation of unfolded proteins in the ER. Furthermore, It has also been examined for tissue distribution(pronounced at the fat body), hormonal regulation (juvenile hormone, insulin and juvenile +transferrin; however, it is not effected by transferrin alone), and the effect of exogenous bacteria (peak at 16 h after infection) on the bPDI mRNA expression. The results suggest that bPDI is a member of the ER stress protein group, and it may play an important role in exogenous bacterial infection in fat body, and that homones regulate its expression.

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Immunogold studies on the gonadotropes in adenohypophysis of the Korean native goat (Immunogold법에 의한 한국재래산양 샘뇌하수체의 성샘자극세포에 관한 연구)

  • Lee, In-se;Lee, Heungshik S.;Song, Seung-hoon;Yoon, Sung-tae;Hwang, In-koo;Seo, Je-hoon;Kang, Tae-cheon;Won, Moo-ho
    • Korean Journal of Veterinary Research
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    • v.39 no.5
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    • pp.921-929
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    • 1999
  • There have been a number of studies of gonadotropes secreting LH and FSH in the adenohypophysis, but the pattern of hormone storage and secretion of these cells still remains a controversial matter. In this study, we examined whether gonadotropes contained both of LH and FSH, and if so, how these hormones were distributed within the secretory granules. Hypophyseal sections of Korean native goat were simultaneously immunostained for LH and FSH antisera by protein A-gold technique. It was found that most gonadotropes contained both FSH and LH, but hormone storages in the secretory granules were some different among cells. Three types of gonadotropes were identified by the shape and size of the secretory granules and their hormone storage patterns. One type(I) of gonadotropes contained oval secretory granules, which immunoreactivity for FSH and LH were very weak. The size of secretory granules ranged from 160 to 310nm in diameter. Most granules contained both FSH and LH, but some contained only one of them. In another type(II) of gonadotropes, the immunreactivity and hormone storage patterns of the secretory granules were similar to those of type I cells. However, the secretory granules were round in shape and larger in size than those of type I. The other gonadotropes(type III) were distinctly distinguished by plenty of hormones in their secretory granules which were densely packed with numerous immunolabelled gold particles. These data are some inconsistent with other results that have been obtained in other ruminants like as cattle and sheep.

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Immunoblot findings of calcareous corpuscles binding proteins in cyst fluid of Taenia solium metacestodes

  • Yang, Hyun-Jong
    • Parasites, Hosts and Diseases
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    • v.42 no.3
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    • pp.141-143
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    • 2004
  • After collecting calcareous corpuscles from plerocercoid of Spirometra mansoni (sparganum), we evaluated the antigenic values of calcareous corpuscles binding proteins obtained from the cyst fluid of Taenia solium metacestodes. Immunoblot analysis revealed that cysticercosis patient sera strongly recognized 10 and 95 kDa calcareous corpuscles binding proteins. This result demonstrated that calcareous corpuscles are bound with major secretory antigenic proteins, which is possibly involved in the secretory pathways of the 10 and 95 kDa proteins presenting in the cyst fluid of T. solium metacestodes.

Multiple Signaling Pathways Contribute to the Thrombin-induced Secretory Phenotype in Vascular Smooth Muscle Cells

  • Jeong, Ji Young;Son, Younghae;Kim, Bo-Young;Eo, Seong-Kug;Rhim, Byung-Yong;Kim, Koanhoi
    • The Korean Journal of Physiology and Pharmacology
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    • v.19 no.6
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    • pp.549-555
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    • 2015
  • We attempted to investigate molecular mechanisms underlying phenotypic change of vascular smooth muscle cells (VSMCs) by determining signaling molecules involved in chemokine production. Treatment of human aortic smooth muscle cells (HAoSMCs) with thrombin resulted not only in elevated transcription of the (C-C motif) ligand 11 (CCL11) gene but also in enhanced secretion of CCL11 protein. Co-treatment of HAoSMCs with GF109230X, an inhibitor of protein kinase C, or GW5074, an inhibitor of Raf-1 kinase, caused inhibition of ERK1/2 phosphorylation and significantly attenuated expression of CCL11 at transcriptional and protein levels induced by thrombin. Both Akt phosphorylation and CCL11 expression induced by thrombin were attenuated in the presence of pertussis toxin (PTX), an inhibitor of Gi protein-coupled receptor, or LY294002, a PI3K inhibitor. In addition, thrombin-induced production of CCL11 was significantly attenuated by pharmacological inhibition of Akt or MEK which phosphorylates ERK1/2. These results indicate that thrombin is likely to promote expression of CCL11 via PKC/Raf-1/ERK1/2 and PTX-sensitive protease-activated receptors /PI3K/Akt pathways in HAoSMCs. We propose that multiple signaling pathways are involved in change of VSMCs to a secretory phenotype.