• Title/Summary/Keyword: adhesive protein

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Total saponin from Korean Red Ginseng inhibits binding of adhesive proteins to glycoprotein IIb/IIIa via phosphorylation of VASP (Ser157) and dephosphorylation of PI3K and Akt

  • Kwon, Hyuk-Woo;Shin, Jung-Hae;Cho, Hyun-Jeong;Rhee, Man Hee;Park, Hwa-Jin
    • Journal of Ginseng Research
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    • v.40 no.1
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    • pp.76-85
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    • 2016
  • Background: Binding of adhesive proteins (i.e., fibrinogen, fibronectin, vitronectin) to platelet integrin glycoprotein IIb/IIIa (${\alpha}IIb/{\beta}3$) by various agonists (thrombin, collagen, adenosine diphosphate) involve in strength of thrombus. This study was carried out to evaluate the antiplatelet effect of total saponin from Korean Red Ginseng (KRG-TS) by investigating whether KRG-TS inhibits thrombin-induced binding of fibrinogen and fibronectin to ${\alpha}IIb/{\beta}3$. Methods: We investigated the effect of KRG-TS on phosphorylation of vasodilator-stimulated phosphoprotein (VASP) and dephosphorylation of phosphatidylinositol 3-kinase (PI3K) and Akt, affecting binding of fibrinogen and fibronectin to ${\alpha}IIb/{\beta}3$, and clot retraction. Results: KRG-TS had an antiplatelet effect by inhibiting the binding of fibrinogen and fibronectin to ${\alpha}IIb/{\beta}3$ via phosphorylation of VASP ($Ser^{157}$), and dephosphorylation of PI3K and Akt on thrombin-induced platelet aggregation. Moreover, A-kinase inhibitor Rp-8-Br-cyclic adenosine monophosphates (cAMPs) reduced KRG-TS-increased VASP ($Ser^{157}$) phosphorylation, and increased KRG-TS-inhibited fibrinogen-, and fibronectin-binding to ${\alpha}IIb/{\beta}3$. These findings indicate that KRG-TS interferes with the binding of fibrinogen and fibronectin to ${\alpha}IIb/{\beta}3$ via cAMP-dependent phosphorylation of VASP ($Ser^{157}$). In addition, KRG-TS decreased the rate of clot retraction, reflecting inhibition of ${\alpha}IIb/{\beta}3$ activation. In this study, we clarified ginsenoside Ro (G-Ro) in KRG-TS inhibited thrombin-induced platelet aggregation via both inhibition of $[Ca^{2+}]_i$ mobilization and increase of cAMP production. Conclusion: These results strongly indicate that KRG-TS is a beneficial herbal substance inhibiting fibrinogen-, and fibronectin-binding to ${\alpha}IIb/{\beta}3$, and clot retraction, and may prevent platelet ${\alpha}IIb/{\beta}3$-mediated thrombotic disease. In addition, we demonstrate that G-Ro is a novel compound with antiplatelet characteristics of KRG-TS.

Screening of Lactic Acid Bacteria with Potent Adhesive Property in Human Colon using Colonic Mucin-binding Assay (Colonic mucin-binding assay를 이용한 장내 우수 점착능 유산균주의 선별)

  • Kim, Seong-Yeong;Shin, Kwang-Soon;Lee, Ho
    • Korean Journal of Food Science and Technology
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    • v.36 no.6
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    • pp.959-967
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    • 2004
  • To screen probiotic lactic acid bacteria with potent adhesive property on human colonic mucosa, colonic mucin-binding assay was introduced. This colonic mucin-binding assay actually measures the binding activity of surface lectin-like protein (SLP) on colonic mucin, and the optimal conditions were examined. The optimal pH for colonic mucin coating on plate wells was 4.8, and ${\times}24,000$ diluted solution of commercially available horseradish peroxidase (HRP) conjugated streptoavidin yielded good results, for rapid screening, $5.0\;{\mu}g/mL$ of biotinylated SLP from lactic acid bacteria was optimal, and optimal scintillation time of 3,3',5,5'-tetramethyl benzidine (TMB) was 10 min. These conditions were useful for both rapid selection and quantitative analysis of lactic acid bacteria that have high adhesion property to human intestinal tract. Among 50 strains of lactic acid bacteria, including 32 type culture strains and 18 isolated strains from infant feces, Lactobacillus species FSB-1 isolated from kimchi showed the highest binding activity to colonic mucin. From taxonomical viewpoints based on morphological study, physico-biochemical study, partial 16S rDNA seguencing, and phylogenetic analysis, L. species FSB-1 was identified as Lactobacillus brevis.

Regulation of an Outer Membrane Protein, OmpW, Expression and Its Biological Function in Salmonell typhimurium (Salmonella typhimurium 외막 단백질 OmpW의 발현조절 및 기능에 관한 연구)

  • Yoo, Ah-Young;Yu, Jong-Earn;Yang, Ji-Seon;Kim, Young-Hee;Bae, Chang-Ho;Oh, Jeong-Il;Kang, Ho-Young
    • Journal of Life Science
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    • v.18 no.11
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    • pp.1606-1611
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    • 2008
  • Outer membrane proteins (OMPs) expressed in the Gram negative bacteria such as Salmonella play multiple functions including material transports, adhesive factors and reception of external signals. This study has been focused on an OmpW protein known as a protein required to form a hydrophobic porin in outer membrane. We have constructed a S. typhimurium CK10 mutant deleting an ompW gene on chromosome. The CK10 strain was more tolerant to SDS than the wild-type strain did. As increase of salt concentration in the culture media, significantly decreased amount of OmpW protein in cells were detected. The maximum OmpW protein was expressed in the absence of salt supplement. However, the growth of CK10 strain was indistinguishable compared to that of the wild-type strain at the variable osmotic conditions. The biological role of differential OmpW expression in response to osmotic conditions remains to be investigated.

Encystation of Giardia lamblia by High Bile and Alkaline pH and Its Ultrastructural Changes during Encystation

  • Yong, Tai-Soon;Yang, Hye-Won;Im, Kyung-Il;Park, Soon-Jung
    • Journal of Microbiology and Biotechnology
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    • v.10 no.3
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    • pp.429-433
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    • 2000
  • Giardia lamblia, a human pathogen causing outbreaks of diarrhea, recently became a focus of great concerns in the fields of both medical and environmental microbioloty. To develop the experimental tools to study giardiasis, encystation, one of the major processes in its life cycle, was reconstituted by inducing an axenic culture of a flagellated form of G. lamblia into a cyst from under high concentration of bile and alkaline pH condition. The successful induction was confirmed by Northern analysis of resulting increased expression of the CWPl gene encoding the cyst wall protein 1. An examination of the encystation process with SEM (scanning electron microscopy) and TEM (transmission electron microscopy) revealed that the trophozoite, a flagellate with a bilateral symmetry, was transformed to a cyst form with an oval-shape and defined filamentous wall. The encystation was found to cause a disappearance of the flagella and an invagination of the adhesive disc. An extensive formation of rER (rough endoplasmic reticulum) was observed after 24h of induction, indication an active synthesis and export of proteins during this process. The vital staining of the invitro-induced systs showed that most cysts maintained their viability.

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Three-Dimensional Visualization for Chemical Components within Rice Particle With a Microslicer-Infrared Microscope System

  • Do, Gab-Soo;Kudoh, Ken-Ichi;Furushiro, Naomichi;Koyama, Ryo;Higuchi, Toshiro;Sagara, Yasuyuki
    • Proceedings of the Korean Society of Near Infrared Spectroscopy Conference
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    • 2001.06a
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    • pp.1531-1531
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    • 2001
  • A novel technique has been developed to observe the three-dimensional (3-D) distribution of chemical components in biological materials using both automatic sectioning microtome and infrared microscope. The 3-D image was reconstructed based on the relationship between the content and the absorbancy of specified wavelength for chemical components. By using the automatic sectioning microtome, the kernel of rice sample fixed in paraffin was sequentially sliced with the thickness of $5\;\mutextrm{m}$ after pasting the sliced sectional specimens on an adhesive tape. The chemical components of the specimens, which are placed on the X-Y controlling stage with positioning accuracy of ${\pm}10\;\mutextrm{m}$, were analyzed by the infrared microscope. The 3-D images demonstrated that the zonal protein about $200\mutextrm{m}$ in width was observed mainly at the outer parts of a rice particle, and carbohydrates entirely. These images can be observed by choosing arbitrary observation angle. The result indicated that the developed technique could be applied 3-D information to investigate intrinsic chemical components but also residual pesticides as well as bacteria contamination for agricultural produce.

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Bio-Inspired Synthesis of a Silicate/PMMA Composite

  • Nam, Kyung Mok;Lee, Yoon Joo;Kwon, Woo Teck;Kim, Soo Ryong;Shin, Dong-Geun;Lim, Hyung Mi;Kim, Hyungsun;Kim, Younghee
    • Journal of the Korean Ceramic Society
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    • v.51 no.1
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    • pp.7-10
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    • 2014
  • Abalone shell is composed of 95 wt% $CaCO_3$ platelets and 5 wt% of a protein-rich organic matrix which acts as an adhesive layer, connecting aragonite tablets, thus maintaining the structural integrity of the composite. By mimicking abalone shell, we prepared a silicate plate/polymer nanocomposite by infiltrating PMMA between silicate layers and warm-pressing them at $200^{\circ}C$ for 1 h under 15 tons to make organic-inorganic composite materials. To examine the organic-inorganic composite materials after the warm-pressing procedure, the composite sample was analyzed with FE-SEM and TG. The bending strengths and densities of the composites prepared by a silicate plate and PMMA after the warm-pressing process were ~140 MPa and 1.5, respectively.

Analysis of the Potent Platelet Glycoprotein IIb-IIIa Antagonist from Natural Sources

  • Kang, In-Cheol;Kim, Doo-Sik
    • BMB Reports
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    • v.31 no.5
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    • pp.515-518
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    • 1998
  • Adhesive interaction of the platelet glycoprotien IIb-IIIa (GP IIb-IIIa) with a plasma protein, such as fibrinogen, plays an important role in thrombosis and hemostasis. The specific sequence Arg-Gly-Asp (RGD) is critical for the binding of fibrinogen to platelet. To examine and characterize the GP IIb-IIIa antagonist from natural sources, we have developed a simple enzyme-linked immunosorbant assay (ELISA) system. The GP IIb-IIIa complex was purified to homogeneity from platelet Iysates by the combination of two affinity chromatographic methods using the synthetic RGD peptide (GRGDSPK)-immobilized Sepharose and wheat germ lectin-Sepharose. The synthetic peptide GRGDSP inhibits GP IIb-IIIa binding to immobilized fibrinogen with an $IC_{50}$ of $1.5\;{\mu}M$. Venoms of three different snake species and a Korean scolopendra extract have strong antagonistic activities for the binding of human fibrinogen to the platelet GP IIb-IIIa complex. The $IC_{50}$ values of the snake venom s and scolopendra were in the range of $5.5\;{\mu}g$ to $60\;{\mu}g$. These results provide meaningful information for developing antiplatelet agents.

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Expression and Clinical Significance of Osteopontin in Calcified Breast Tissue

  • Huan, Jin-Liang;Xing, Li;Qin, Xian-Ju;Gao, Zhi-Guang;Pan, Xiao-Feng;Zhao, Zhi-Dong
    • Asian Pacific Journal of Cancer Prevention
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    • v.13 no.10
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    • pp.5219-5223
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    • 2012
  • Osteopontin (OPN) is an integrin-binding protein, believed to be involved in a variety of physiological cellular functions. The physiology of OPN is best documented in the bone where this secreted adhesive glycoprotein appears to be involved in osteoblast differentiation and bone formation. In our study, we used semi-quantitative RT-PCR of osteopontin in calcification tissue of breast to detect breast cancer metastasis. The obtained data indicate that the expression of osteopontin is related to calcification tissue of breast, and possibly with the incidence of breast cancer. The expression strength of OPN by RT-PCR detection was related to the degree of malignancy of breast lesions, suggesting a close relationship between OPN and breast calcification tissue. The results revealed that expression of OPN mRNA is related to calcification of breast cancer tissue and to the development of breast cancer. Determination of OPN mRNA expression can be expected to be a guide to clinical therapy and prediction of the prognosis of breast cancer patients.

Expression of Ajuba, a Novel LIM Protein, is Regulated by Endorlasmic Reticulum Stress (소포체 스트레스가 Ajuba 발현유도)

  • Park, Sang-Mi;Kwon, Ki-Sang;Yun, Eun-Young;Goo, Tae-Won;Kwon, O-Yu
    • Journal of Life Science
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    • v.17 no.7 s.87
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    • pp.1023-1025
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    • 2007
  • Ajuba is a number of proteins containing cytosolic LIM domain. Its function may provide a new pathway whereby cell-cell adhesive events are transmitted to the nucleus to regulate cell proliferation and differentiation decisions. Here, Ajuba gene expression was investigated its molecular properties associated with endoplasmic reticulum (ER) stresses (tunicamycin, DTT, A23187 and BFA) which induced remarkable ex-pression of Ajuba mRNA. The mRNA half life of Ajuba was also determined, its half life of Ajuba mRNA in FRTL-5 cells was approximately 2 hr after the initial translation. Although the obvious bioligical function of Ajuba is not clear, on the base of the results, Ajuba gene expression is deeply associated with ER stresses.

THE EFFECT OF THE REMOVAL OF CHONDROITIN SULFATE ON BOND STRENGTH OF DENTIN ADHESIVES AND COLLAGEN ARCHITECTURE (비교원성 단백질이 상아질 접착제의 결합강도와 교원질의 형태에 미치는 영향)

  • Kim, Jong-Ryul;Park, Sang-Jin;Choi, Gi-Woon;Choi, Kyoung-Kyu
    • Restorative Dentistry and Endodontics
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    • v.35 no.3
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    • pp.211-221
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    • 2010
  • Proteoglycan is highly hydrophilic and negatively charged which enable them attract the water. The objective of study was to investigate the effects of Proteoglycan on microtensile bond strength of dentin adhesives and on architecture of dentin collagen matrix of acid etched dentin by removing the chondroitin sulphate attached on Proteoglycan. A flat dentin surface in mid-coronal portion of tooth was prepared. After acid etching, half of the specimens were immersed in 0.1 U/mL chondroitinase ABC (C-ABC) for 48 h at $37^{\circ}C$, while the other half were stored in distilled water. Specimens were bonded with the dentin adhesive using three different bonding techniques (wet, dry and re-wet) followed by microtensile bond strength test. SEM examination was done with debonded specimen, resin-dentin interface and acid-etched dentin surface with/without C-ABC treatment. For the subgroups using wet-bonding or dry-bonding technique, microtensile bond strength showed no significant difference after C-ABC treatment (p > 0.05). Nevertheless, the subgroup using rewetting technique after air dry in the Single Bond 2 group demonstrated a significant decrease of microtensile bond strength after C-ABC treatment. Collagen architecture is loosely packed and some fibrils are aggregated together and relatively collapsed compared with normal acid-etched wet dentin after C-ABC treatment. Further studies are necessary for the contribution to the collagen architecture of noncollagenous protein under the various clinical situations and several dentin conditioners and are also needed about long-term effect on bond strength of dentin adhesive.