• Title/Summary/Keyword: Endoplasmic reticulum (ER)

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Inhibition of Store-Operated Calcium Entry Protects Endothelial Progenitor Cells from H2O2-Induced Apoptosis

  • Wang, Yan-Wei;Zhang, Ji-Hang;Yu, Yang;Yu, Jie;Huang, Lan
    • Biomolecules & Therapeutics
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    • v.24 no.4
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    • pp.371-379
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    • 2016
  • Store-operated calcium entry (SOCE), a major mode of extracellular calcium entry, plays roles in a variety of cell activities. Accumulating evidence indicates that the intracellular calcium ion concentration and calcium signaling are critical for the responses induced by oxidative stress. The present study was designed to investigate the potential effect of SOCE inhibition on $H_2O_2$-induced apoptosis in endothelial progenitor cells (EPCs), which are the predominant cells involved in endothelial repair. The results showed that $H_2O_2$-induced EPC apoptosis was reversed by SOCE inhibition induced either using the SOCE antagonist ML-9 or via silencing of stromal interaction molecule 1 (STIM1), a component of SOCE. Furthermore, SOCE inhibition repressed the increases in intracellular reactive oxygen species (ROS) levels and endoplasmic reticulum (ER) stress and ameliorated the mitochondrial dysfunction caused by $H_2O_2$. Our findings provide evidence that SOCE inhibition exerts a protective effect on EPCs in response to oxidative stress induced by $H_2O_2$ and may serve as a potential therapeutic strategy against vascular endothelial injury.

Fyn Kinase: A Potential Therapeutic Target in Acute Kidney Injury

  • Uddin, Md Jamal;Dorotea, Debra;Pak, Eun Seon;Ha, Hunjoo
    • Biomolecules & Therapeutics
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    • v.28 no.3
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    • pp.213-221
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    • 2020
  • Acute kidney injury (AKI) is a common disease with a complex pathophysiology which significantly contributes to the development of chronic kidney disease and end stage kidney failure. Preventing AKI can consequently reduce mortality, morbidity, and healthcare burden. However, there are no effective drugs in use for either prevention or treatment of AKI. Developing therapeutic agents with pleiotropic effects covering multiple pathophysiological pathways are likely to be more effective in attenuating AKI. Fyn, a non-receptor tyrosine kinase, has been acknowledged to integrate multiple injurious stimuli in the kidney. Limited studies have shown increased Fyn transcription level and activation under experimental AKI. Activated Fyn kinase propagates various downstream signaling pathways associated to the progression of AKI, such as oxidative stress, inflammation, endoplasmic reticulum stress, as well as autophagy dysfunction. The versatility of Fyn kinase in mediating various pathophysiological pathways suggests that its inhibition can be a potential strategy in attenuating AKI.

Expression of Lymphocyte ADP-ribosyltransferase in Rat Mammary Adenocarcinoma Cells (임파구 ADP-ribosyltransferase의 rat mammary adenocarcinoma cell에서의 발현)

  • 김현주
    • Journal of Life Science
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    • v.8 no.1
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    • pp.102-108
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    • 1998
  • The nascent from of glycosylphosphatidylinositol (GPI)-anchored proteins possesses both amino and carboxy terminal hydrophobic signal sequences to direct processing in the endoplasmic reticulum (ER). Following cleavage of the amino-terminal signal peptide, the carboxy-terminal peptide is processed. Previously, mouse lymphocyte NDA: agrinine ADP-ribosyltransferase (Yac-1) was cloned and the deduced amino acid sequence of the Yac-1 transferase contained hydrophobic amino and carboxy termini, consistent with known signal sequences of GPI-anchored proteins. This tranferase was present on the surface of NMU (rat mammary adenocarcinoma) cells transfected with the wildtype cDNA and was released with phosphatidylinositol-specific phosphilpase C. Expression of the mutant protein, lacking the carboxy terminal hydrophobic sequence, resulted in the peoduction of soluble, secreted from of the transferase. This result shows that carboxy terminal sequence is important for GPI-attachment.

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Assay of the Bombyx mori Recombinant Protein Disulfide Isomerase (bPDI) Acivity

  • Goo, Tae-Won;Yun, Eun-Young;Kim, Sung-Wan;Park, Kang-Ho;Hwang, Jae-Sam;Kang, Seok-Woo;Park, Soo-Jung;Kwon, O-Yu
    • Proceedings of the Korean Society of Sericultural Science Conference
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    • 2003.04a
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    • pp.45-45
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    • 2003
  • Protein disulfide isomerase (PDO) is an essential protein which is localized to the endoplasmic reticulum (ER) of eukaryotic cells. It catalyses the formation and isomerization of disulfide bonds during the folding of secretory proteins. We have isolarted a cDNA encoding PDI from Bombyx mori (bPDI), in which an open reading frame of 494 mino acid (55.6kDa) is shown. (omitted)

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ASIC2a-dependent increase of ASIC3 surface expression enhances the sustained component of the currents

  • Kweon, Hae-Jin;Cho, Jin-Hwa;Jang, Il-Sung;Suh, Byung-Chang
    • BMB Reports
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    • v.49 no.10
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    • pp.542-547
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    • 2016
  • Acid-sensing ion channels (ASICs) are proton-gated cation channels widely expressed in the nervous system. Proton sensing by ASICs has been known to mediate pain, mechanosensation, taste transduction, learning and memory, and fear. In this study, we investigated the differential subcellular localization of ASIC2a and ASIC3 in heterologous expression systems. While ASIC2a targeted the cell surface itself, ASIC3 was mostly accumulated in the ER with partial expression in the plasma membrane. However, when ASIC3 was co-expressed with ASIC2a, its surface expression was markedly increased. By using bimolecular fluorescence complementation (BiFC) assay, we confirmed the heteromeric association between ASIC2a and ASIC3 subunits. In addition, we observed that the ASIC2a-dependent surface trafficking of ASIC3 remarkably enhanced the sustained component of the currents. Our study demonstrates that ASIC2a can increase the membrane conductance sensitivity to protons by facilitating the surface expression of ASIC3 through herteromeric assembly.

Expression of Recombinant Human Stem Cell Factor (hSCF) Protein using Bombyx mori Protein Disulfide Isomerase (bPDI)

  • Kim, Sung-Wan;Yun, Eun-Young;Kim, Seong-Ryul;Park, Seung-Won;Kang, Seok-Woo;Lee, Kwang-Gill;Kwon, O-Yu;Goo, Tae-Won
    • International Journal of Industrial Entomology and Biomaterials
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    • v.21 no.2
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    • pp.151-155
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    • 2010
  • Protein disulfide isomerase (PDI) catalyzes the oxidation of disulfides and the isomerizatiob of incorrect disulfides in new polypeptides during folding in the oxidizing environment of the endoplasmic reticulum (ER). To increase recombinant protein hSCF (human stem cell factor) production, we have developed expression system using the Bombyx mori PDI (bPDI) as a fusion partner. bPDI gene fusion was found to improve the production of recombinant hSCFs. Thus, we conclude that bPDI gene fusion will be very useful for the large-scale production of biologically active recombinant proteins.

Expression of Aβ-Fc Fusion Protein in Transgenic Potato

  • Kim, Hyun-Soon;Youm, Jung Won;Lee, Jeong-Hwan;Jeon, Jae-Heung;Ko, Kisung
    • Horticultural Science & Technology
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    • v.32 no.3
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    • pp.375-381
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    • 2014
  • Transgenic potato was generated to express recombinant 5 repeated ${\beta}$-amyloid ($A{\beta}$) peptides, potential antigens to be applied as a preventive accine for Alzheimer's disease using Agrobacterium mediated transformation. The $A{\beta}$ peptides were fused to the human IgG Fc fragment enhancing protein and KDEL, which is the endoplasmic reticulum (ER) retention signal ($5A{\beta}$-FcK). The $5A{\beta}$-FcK, was expressed under the control of the duplicated 35S promoter. PCR analysis confirmed the presence of the transgene in several transgenic potato lines. Southern blot analysis showed only a single gene copy number in transgenic line 22, whereas multiple gene copy numbers were shown for transgenic lines 31 and 44. Northern blot analysis showed that line 22 had stronger mRNA levels when compared to lines 31 and 44. Immunoblot analysis confirmed that the $5A{\beta}$-FcK protein was expressed in the transgenic potato plant. These results indicate that $5A{\beta}$ fused to Fc can be expressed in potato plants.

Effects of Cyclophosphamide in the Epididymis of the Rat II. Corpus (Cyclophosphamide가 흰쥐의 부정소에 미치는 영향 II. 체 부)

  • Cho, Kwang-Phil;Kim, Jeong-Sang;Jung, Hae-Man
    • Applied Microscopy
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    • v.22 no.2
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    • pp.127-140
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    • 1992
  • This research was undertaken to determine the effects of the cyclophosphamide (CP) on the epididymal corpus of the male rat in terms of ultrastructural alteration and protein analysis by SDS-PAGE at different groups; control group, 1 week group, 3 weeks group and 5 weeks group were treated with saline (control group) or CP at doses of 20mg/kg/week, 1 time a week, respectively. In the cytoplasm of the principal cells on the epididymal corpus, the mitochondria were significantly swollen or disrupted. The lumens of rough endoplasmic reticulum (rER) were also dilated and the number of secretory vesicles and lysosomes were increased respectively. CP caused changes in protein concentrations in the corpus of epididymis after CP treatment. Total proteins of 31 to 36 species were expressed in the corpus fluid. Then the more CP was increased, the more concentration of proteins caused to decrease, synthesize or increase in epididymal corpus. In contrast to the control group, in particular 88KD and the other 8 proteins in the corpus fluid, were decreased or disappeared respectively, whereas acid phosphatase and the other 9 proteins in the corpus fluid, were increased or synthesized respectively. The other proteins are not showed distinctive difference. It is suggested that treatment with CP alters the specific cell organelles and proteins in segment of the epididymal corpus.

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Upregulation of Kruppel-like Factor 4 Gene expression by Allomyrina dichotoma Hemolymph in the INS-1 Pancreatic β-cells

  • Kwon, Kisang;Suh, Hyun-Woo;Kim, Hong Geun;Kwon, O-Yu
    • Biomedical Science Letters
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    • v.26 no.1
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    • pp.37-41
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    • 2020
  • The hemolymph of Korean rhinoceros Allomyrina dichotoma consists of blood and lymph in which various kinds of proteins function physiologically. We have previously demonstrated that A. dichotoma hemolymph has the potential to treatment and prevent diabetes through activating transcription factor 3-gene (ATF3) regulation. In this study, we investigate the expression of Kruppel-like factor 4 (KLF4) in A. dichotoma hemolymph-treated INS-1 pancreatic β-cells. The new findings show that A. dichotoma hemolymph, which upregulates KLF4 gene expression in a dose-dependent and time-dependent manner. In addition, hemolymph combine with mild endoplasmic reticulum (ER) stress, which also differentially regulates KLF4 gene expression. These results may provide insights to KLF4 gene-related disease therapies through KLF4 gene regulation.