• Title/Summary/Keyword: Dense granule protein

Search Result 14, Processing Time 0.023 seconds

Effects of specific monoclonal antibodies to dense granular proteins on the invasion of Toxoplasma gondii in vitro and in vivo

  • Cha, Dong-Yeob;Song, In-Kwan;Lee, Gye-Sung;Hwang, Ok-Sun;Noh, HyungJun;Yeo, Seung-Dong;Shin, Dae-Whan;Lee, Young-Ha
    • Parasites, Hosts and Diseases
    • /
    • v.39 no.3
    • /
    • pp.233-240
    • /
    • 2001
  • Although some reports have been published on the protective effect of antibodies to Toxoplasma gondii surface membrane proteins, few address the inhibitory activity of antibodies to dense granular proteins (GRA proteins) . Therefore, we performed a series of experiments to evaluate the inhibitory effects of monoclonal antibodies (mAbs) to GRA proteins (GRA2, 28 kDa; GRA6, 32 kDa) and surface membrane protein (SAGI, 30 kDa) on the invasion of T. gondii tachyzoites. Passive immunization of mice with one of three mAbs following challenge with a lethal dose of tachyzoites significantly increased survival compared with results for mice treated with control ascites. The survival times of mice challenged with tachyzoties pretreated with anti-GRA6 or anti-SAG 1 mAb were significantly increased. Mice that received tachyzoties pretreated with both mAb and complement had longer survival times than those that received tachyzoites pretreated with mAb alone. Invasion of tachyzoites into fibroblasts and macrophages was significantly inhibited in the anti-GRA2, anti-GRA6 or anti-SAG 1 mAb pretreated group. Pretreatment with mAb and complement inhibited invasion of tachyzoites in both fibroblasts and macrophages. These results suggest that specific antibodies to dense-granule molecules may be useful for controlling infection with T. gondii.

  • PDF

Ultrastructural characteristics of the pancreatic acinar cells in the diabetes-prone BB rat (BB랫드의 췌장 샘포세포의 미세구조적 특징)

  • Jeong, Young-gil;Kim, Kil-soo;Lee, Chul-ho;Won, Moo-ho;Cho, Sung-whan;Kim, Moo-kang
    • Korean Journal of Veterinary Research
    • /
    • v.36 no.1
    • /
    • pp.1-9
    • /
    • 1996
  • Functional and morphological characteristics of the exocrine pancreas in genetic model BB rat of insulin dependent diabetes medllitus(IDDM) were carried out. Wistar rat was used as control animal. Flow rate of pancreatic juice, output of amylase and protein, and plasma glucose and insulin levess were examined. Also light and ultrastructural characteristics of the exocrine pancreas were observed. Pancreatic flow rate, output of amylase and protein, and insulin level were lower;glucose level was higher comparing with those of the control Wistar rat. In Wistar rat, exocrine pancreas was typical light microscopically. Zymogen granules and cell organelles were well developed in fine structure. Cell size of the periinsular acini was larger, and number of zymogen granules were more than those of the teleinsular acini. Most acinar cells were dark cells which containe well-developed RER in their cytoplasm. On the other hand, some light cells which have the dilated RER cisterns were found. In BB rat exocrine pancreas, cell size of per-and tele-insular acini similar to that of Wistar rat. The number of light cells occupied 40-50% compairing with that of Wistar rat. Zymogen granules were lower in number than that of Wistar rat and divied into three types in morphological characteristics ; type I showing normal structure, type II showing the wide hallo and small electron dense core in center of the zymogen granule and type III not having the electron dense core in the zymogen granule. The present ratio of type I, type II and type III are less than 5%, 30-40% and more than 50%, respectively.

  • PDF

Immunocytochemical Localization of Storage Protein in Pea (Pisum sativum) Cotyledon

  • Yu, Seong-Cheol;Lee, Chang-Seob;Kim, Woo-Kap
    • Journal of Plant Biology
    • /
    • v.39 no.2
    • /
    • pp.123-126
    • /
    • 1996
  • The pattern of seed storage protein, vicilin, deposition and site of intracellular localization was examined in cotyledon cells of pea (Pisum sativum) seed using the immunocytochemical methods. The vicilin was confined to the cisternae fo the rough endoplasmic reticulum and dictyosome as well as protein granules newly formed in rough endoplasmic reticulum. Vacuolar protein deposites and protein bodies were also labelled by gold particles. After small protein bodies were formed in the rough endoplasmic reticulum, they were transported to large protein bodies and then fused together. Electron dense protein granule, elaborated in the dictyosome, appears to be transported from dictyosome to protein body. A few unlabelled protein granules seem to be accumulated in other type of proteins than vicilin.

  • PDF

Comprehensive Proteome Analysis of the Excretory/Secretory Proteins of Toxoplasma gondii

  • Lee, Won-Kyu;Ahn, Hye-Jin;Baek, Je-Hyun;Lee, Chong-Heon;Yu, Yeon Gyu;Nam, Ho-Woo
    • Bulletin of the Korean Chemical Society
    • /
    • v.35 no.10
    • /
    • pp.3071-3076
    • /
    • 2014
  • Proteomic analyses of the excretory/secretory proteins from the RH strain of Toxoplasma gondii have been performed to understand their functions in the host-parasite interaction. A total of 34 proteins were identified from LC/MS/MS analysis and their abundance was estimated by spectral counting methods. Among them, 8 species of micronemal proteins (MICs), 2 species of rhoptry proteins (ROPs), and 6 species of dense granular proteins (GRAs) were confirmed. Besides these, 18 species of protein were newly identified, and their cellular functions were estimated from sequence analysis. The three most abundant of the 34 identified extractor/secretory proteins-GRA1, GRA7 and GRA2-were confirmed to be highly expressed in T. gondii using the spectral count method. This phenomenon is another demonstration of the importance of GRA proteins for the penetration and survival of T. gondii.

Immunohistochemical studies of glucagon and somatostatin in the pancreas of the Korean tree squirrel. Sciurus vlugar is corea (청설모췌장의 glucageon과 somatostatin 세포의 면역조직학적 연구)

  • Lee, Hyeung-sik;Lee, Jae-hyun
    • Korean Journal of Veterinary Research
    • /
    • v.33 no.4
    • /
    • pp.573-577
    • /
    • 1993
  • The pancreatic endocrine cells, glucagon and somatostation, of the Korean tree squirrel. Sciurus vulgais corea, were investigation by means of light and electron microscopic immunohistochemistry using the PAP and protein A-gold techniques. Glucagon-immunoreactive cells were distributed the periphery and occasinonaly central region of the pancreatic islets. Also, isolated cell was found between the pancreatic ancinar cells. The glucagon cells contraine granules with a diameter of 240~320nm and the electron dense core usually surrounded by a halo of less dense granular material. The core of granule was labelled strongly with gold particles. Somatostatin-immunoreactive cells were weakly stained in scattered along the peripheral pancreatic islets and were distributed as singly or small groups with in the pancreatic acinar cells. The somatostatin cells were spherical with a diameter of 250~275nm, moderately electron opaque (Gold particles were mostly demonstrated on the entire granule.

  • PDF

Comparative antiplatelet and antithrombotic effects of red ginseng and fermented red ginseng extracts

  • Irfan, Muhammad;Lee, Yuan Yee;Lee, Ki-Ja;Kim, Sung Dae;Rhee, Man Hee
    • Journal of Ginseng Research
    • /
    • v.46 no.3
    • /
    • pp.387-395
    • /
    • 2022
  • Background: Fermentation may alter the bioavailability of certain compounds, which may affect their efficacy and pharmacological responses. This study investigated the antiplatelet effects of red ginseng extract (RGE) and fermented red ginseng extract (FRG). Methods: A rodent model was used to evaluate the antiplatelet and antithrombotic effects of the extracts. Rats were orally fed with human equivalent doses of the extracts for 1 week and examined for various signaling pathways using standard in vivo and ex vivo techniques. Light transmission aggregometry was performed, and calcium mobilization, dense granule secretion, integrin αIIbβ3-mediated signaling molecules, cyclic nucleotide signaling events, and various protein molecules were evaluated ex vivo in collagen-stimulated washed platelets. Furthermore, antithrombotic properties were evaluated using a standard acute pulmonary thromboembolism model, and the effects on hemostasis were investigated using rat and mice models. Results: Both RGE and FRG significantly inhibited platelet aggregation, calcium mobilization, and dense granule secretion along with integrin-mediated fibrinogen binding and fibrinogen adhesion. cAMP levels were found to be elevated in RGE-treated rat platelets. Ginseng extracts did not exert any effect on prothrombin time and activated partial thromboplastin time. RGE-treated mice showed significantly better survival under thrombosis than FRG-treated mice, with no effects on hemostasis, whereas FRG-treated mice exhibited a slight increment in bleeding time. Conclusion: Both extracts, especially RGE, are remarkable supplements to maintain cardiovascular health and are potential candidates for the treatment and prevention of platelet-related cardiovascular disorders.

A Rapid Diagnostic Test for Toxoplasmosis using Recombinant Antigenic N-terminal Half of SAG1 Linked with Intrinsically Unstructured Domain of GRA2 Protein

  • Song, Kyoung Ju;Yang, Zhaoshou;Chong, Chom-Kyu;Kim, Jin-Soo;Lee, Kyung Chan;Kim, Tong-Soo;Nam, Ho-Woo
    • Parasites, Hosts and Diseases
    • /
    • v.51 no.5
    • /
    • pp.503-510
    • /
    • 2013
  • Toxoplasma gondii is an apicomplexan parasite with a broad host range of most warm-blooded mammals including humans, of which one-thirds of the human population has been infected worldwide which can cause congenital defects, abortion, and neonatal complications. Here, we developed a rapid diagnostic test (RDT) for T. gondii infection. Antigenic N-terminal half of the major surface antigen (SAG1) was linked with intrinsically unstructured domain (IUD) of dense granule protein 2 (GRA2). The recombinant GST-GRA2-SAG1A protein was successfully expressed and purified as 51 kDa of molecular weight. Furthermore, antigenicity and solubility of the rGST-GRA2-SAG1A protein were significantly increased. The overall specificity and sensitivity of GST-GRA2-SAG1A loaded RDT (TgRDT) were estimated as 100% and 97.1% by comparing with ELISA result which uses T. gondii whole cell lysates as the antigen. The TgRDT tested with Uganda people sera for field trial and showed 31.9% of seroprevalence against T. gondii antibody. The TgRDT is proved to be a kit for rapid and easy to use with high accuracy, which would be a suitable serodiagnostic tool for toxoplasmosis.

Ultrastructure of Compound Starch Granules and Protein Bodies of Starchy Endosperm Cell in Rice (쌀 배유세포 전분복합체와 단백질체의 미세구조)

  • Chang, Byung-Soo;Lee, Soo-Jeong;Kim, Sung-Kon
    • Applied Biological Chemistry
    • /
    • v.39 no.5
    • /
    • pp.379-383
    • /
    • 1996
  • The ultrastructure of the compound starch granules and the protein bodies of Odaebyeo rice of early matured variety were examined by light microscope and electron microscope. The endosperm cell appealed rectangular or octangular shape on the cross section. The thickness of cell wall containing of membraneous materials was about $0.5\;{\mu}m$ in diameter. The starch cell was filled compactly with globular or oval shaped compound starch granules with the size of $20{\sim}25\;{\mu}m$ in diameter. The compound starch granules were consisted of central core starch granule and concentrical $2{\sim}3$ layers of starch granules. The average thickness of the starch granules were about $5\;{\mu}m$. Most protein bodies were found in the aleurone layer The globular protin bodies were scattered near the compound starch granules and $2.5{\sim}3\;{\mu}m$ in diameter. The protein bodies composed of central electron dense materials and peripheral electron loose materials in limiting membrane.

  • PDF

Generation and Immunity Testing of a Recombinant Adenovirus Expressing NcSRS2-NcGRA7 Fusion Protein of Bovine Neospora caninum

  • Jia, Li-Jun;Zhang, Shou-Fa;Qian, Nian-Chao;Xuan, Xue-Nan;Yu, Long-Zheng;Zhang, Xue-Mei;Liu, Ming-Ming
    • Parasites, Hosts and Diseases
    • /
    • v.51 no.2
    • /
    • pp.247-253
    • /
    • 2013
  • Neospora caninum is the etiologic agent of bovine neosporosis, which affects the reproductive performance of cattle worldwide. The transmembrane protein, NcSRS2, and dense-granule protein, NcGRA7, were identified as protective antigens based on their ability to induce significant protective immune responses in murine neosporosis models. In the current study, NcSRS2 and NcGRA7 genes were spliced by overlap-extension PCR in a recombinant adenovirus termed Ad5-NcSRS2-NcGRA 7, expressing the NcSRS2-NcGRA7 gene, and the efficacy was evaluated in mice. The results showed that the titer of the recombinant adenovirus was $10^9TCID_{50}/ml$. Three weeks post-boost immunization (w.p.b.i.), the IgG antibody titer in sera was as high as 1:4,096. IFN-${\gamma}$ and IL-4 levels were significantly different from the control group (P<0.01). This research established a solid foundation for the development of a recombinant adenovirus vaccine against bovine N. caninum.

Proteomic Analysis of Toxoplasma gondii KI-1 Tachyzoites

  • Choi, Si-Hwan;Kim, Tae-Yun;Park, Sung-Goo;Cha, Guang-Ho;Shin, Dae-Whan;Chai, Jong-Yil;Lee, Young-Ha
    • Parasites, Hosts and Diseases
    • /
    • v.48 no.3
    • /
    • pp.195-201
    • /
    • 2010
  • We studied on the proteomic characteristics of Toxoplasma gondii KI-1 tachyzoites which were originally isolated from a Korean patient, and compared with those of the well-known virulent RH strain using 2-dimensional electrophoresis (2-DE), mass spectrometry, and quantitative real-time PCR. Two-dimensional separation of the total proteins isolated from KI-1 tachyzoites revealed up to 150 spots, of which 121 were consistent with those of RH tachyzoites. Of the remaining 29 spots, 14 showed greater than 5-fold difference in density between the KI-1 and RH tachyzoites at a pH of 5.0-8.0. Among the 14 spots, 5 from the KI-1 isolate and 7 from the RH strain were identified using MALDI-TOF mass spectrometry and database searches. The spots from the KI-1 tachyzoties were dense granule proteins (GRA 2,3,6, and 7), hypoxanthine-guanine-xanthine phosphoribosyltransferase (HGRPTase), and uracil phosphoribosyltransferase (UPRTase). The spots from the RH strain were surface antigen 1 (SAG 1), L-lactate dehydrogenase (LDH), actin, chorismate synthase, peroximal catalase, hexokinase, bifunctional dihydrofolate reductase-thymidylate synthase (DHTR-TS), and nucleosidetriphosphatases (NTPases). Quantitative real-time PCR supported our mass spectrometric results by showing the elevated expression of the genes encoding GRA 2,3, and 6 and UPRTase in the KI-1 tachyzoites and those encoding GRA 7, SAG 1, NTPase, and chorismate synthase in the RH tachyzoites. These observations demonstrate that the protein compositions of KI-1 and RH tachyzoites are similar but differential protein expression is involved in virulence.