• Title/Summary/Keyword: Protein Transduction

Search Result 599, Processing Time 0.031 seconds

Levosulpiride, (S)-(-)-5-Aminosulfonyl-N-[(1-ethyl-2-pyrrolidinyl) methyl]-2-methoxybenzamide, enhances the transduction efficiency of PEP-1-ribosomal protein S3 in vitro and in vivo

  • Ahn, Eun-Hee;Kim, Dae-Won;Kim, Duk-Soo;Woo, Su-Jung;Kim, Hye-Ri;Kim, Joon;Lim, Soon-Sung;Kang, Tae-Cheon;Kim, Dong-Joon;Suk, Ki-Tae;Park, Jin-Seu;Luo, Qiuxiang;Eum, Won-Sik;Hwang, Hyun-Sook;Choi, Soo-Young
    • BMB Reports
    • /
    • v.44 no.5
    • /
    • pp.329-334
    • /
    • 2011
  • Many proteins with poor transduction efficiency were reported to be delivered to cells by fusion with protein transduction domains (PTDs). In this study, we investigated the effect of levosulpiride on the transduction of PEP-1 ribosomal protein S3 (PEP-1-rpS3), and examined its influence on the stimulation of the therapeutic properties of PEP-1-rpS3. PEP-1-rpS3 transduction into HaCaT human keratinocytes and mouse skin was stimulated by levosulpiride in a manner that did not directly affect the cell viability. Following 12-O-tetradecanoylphorbol-13-acetate (TPA)-induced inflammation in mice, levosulpiride alone was ineffective in reducing TPA-induced edema and in inhibiting the elevated productions of inflammatory mediators and cytokines, such as cyclooxygenase-2, inducible nitric oxide synthase, interleukin-6 and -1${\beta}$, and tumor necrosis factor-${\alpha}$. Anti-inflammatory activity by PEP-1-rpS3 + levosulpiride was significantly more potent than by PEP-1-rpS3 alone. These results suggest that levosulpiride may be useful for enhancing the therapeutic effect of PEP-1-rpS3 against various inflammatory diseases.

A Study for the Mechanism of Abnormal Proliferation in Vascular Endothelial Cells using Inhibitors to the Signal Transduction Pathway (신호전달 경로의 저해제를 이용한 혈관 내피세포의 비정상적인 증식 기전에 대한 연구)

  • Bae, Yong Chan;Park, Suk Young;Nam, Su Bong;Herh, Jae Young;Kang, Young Seok
    • Archives of Plastic Surgery
    • /
    • v.33 no.1
    • /
    • pp.5-12
    • /
    • 2006
  • Protein tyrosine kinase(PTK), protein kinase C(PKC), oxidase, as a mediator, take a significant role in signal transduction pathway of angiogenesis. The authors utilized the inhibitors, targeting the formation of three co-enzyme in signal transduction pathway in order to quantify the suppression of abnormal vascular endothelial cell proliferation induced by DMH, to compare the level suppression in each up-regulated growth factors, CTGF, CYR61, $ITG{\beta}1$, FHL2, and to identify the relationship between abnormal cell proliferation and signal transduction pathway. Five groups were established; Control group, Group of DMH, Group of DMH-mixed Herbimycin, inhibitor of protein tyrosine kinase, Group of DMH-mixed Calphostin C, inhibitor of protein kinase C, Group Of Dmh-Mixed 10U Catalase, Inhibitor Of oxidase. The rise of vascular endothelial cell was compared by MTT assay, and four growth factors were analysed with RT-PCR method, at pre-administration, 4, 8, 12, and 24 hours after administration. In comparison of abnormal proliferation of vascular endothelial cell induced by DMH, suppression was noticed in Herbimycin and Calphostin C group, and Calphostin C group revealed higher suppression effect. Nevertheless, Catalase group did not have any suppression. In manifestation of four growth factors, Herbimycin and Calphostin C group presented similar manifestation with control group, except in $ITG{\beta}$. Catalse group had similar manifestation with DMH group in all four growth factors. Abnormal proliferation of vascular endothelial cell induced by DMH have a direct relationship with PTK and PKC, more specifically to PKC. Oxidase was confirmed not to have any relevance.

G Protein-Coupled Receptors: Molecular Organization and Regulatory Mechanisms

  • Caron, Marc G.
    • Proceedings of the Korean Society of Applied Pharmacology
    • /
    • 1994.04a
    • /
    • pp.82-87
    • /
    • 1994
  • Signal transduction through G protein-coupled receptors comprises three functional components, a receptor, a G protcin and a effector protein. Work over the last sevcral ycars has led to the characterization or virtually all of the components or these systems. what has come out or those studies is that these mechanisms of signal transduction are pervasive in nature being found in mammalian and avian species, as well as lower organisms such as yeast and slime mold. It is known that G protein-coupled receptors mediate the action of such diverse molecules such as small hormones and neurotransmitters, small peptide molecules as well as glycoprotein hormones and various sensory perceptions such as light, olfaction and most likely taste.

  • PDF

Bone regenerative effects of recombinant human bone morphogenetic protein-2 employed protein transduction domain (Protein transduction domain을 이용한 recombinant human bone morphogenetic protein-2의 골재생효과)

  • Jung, Sung-Won;Kim, Nam-Hee;Yook, Jong-In;Kim, Chang-Sung;Kim, Hyung-Jun;Cho, Kyoo-Sung
    • Journal of Periodontal and Implant Science
    • /
    • v.37 no.3
    • /
    • pp.497-509
    • /
    • 2007
  • Bone morphogenetic proteins(BMPs) are regarded as members of the transforming growth $factor-{\beta}$ superfamily with characteristic features in their amino acid sequences. A number of studies have demonstrated the biologic activities of BMPs, which include the induction of cartilage and bone formation. Recently there was a attempt to overcome a limitation of mass production, and economical efficieny of rh-BMPs. The method producing PTD by using bacteria have advantages of acquiry a mass of proteins. Hences, a new treatment which deliver protein employed by protein transduction domain(PTD) has been tried. The purpose of this study was to evaluate the bone regenerative effect of TATBMP-2 and TAT-HA2-BMP-2 employed by PTD from HlV-1 TAT protein for protein translocation in the rat calvarial model. An 8mm calvarial, critical size osteotomy defect was created in each of 32 male Spraque-Dawley rats(weight $250{\sim}300g$). The animals were divided into 4 groups of 32 animals each (4 animals/group/healing interval). The defect was treated with TATBMP-2/ACS(Absorbable collagen sponge) (TATBMP-2 0.1mg/ml), TAT-HA2-BMP-2/ACS(TAT-HA2-BMP-2 0.1mg/ml), ACS alone or left untreated for surgical control(negative control). The rats were sacrificed at 2 or 8 weeks postsurgery, and the results were evaluated histologically. The results were as follows: New bone formation were not significantly greater in the TATBMP-2/ACS group relative to negative, and positive control groups. New bone was evident at the defect sites in TAT-HA2-BMP-2/ACS group relative to negative, positive control and TATBMP-2 groups. There were a little bone regeneration in TATBMP-2 groups. While, enhanced local bone formation were observed in TAT-HA2-BMP-2 group. But, The results was not the same in all rat defects. Therefore, further investigations are required to develop a method. which disperse homogenously, and adhere to target cells.

Nonviral Gene Delivery by a Novel Protein Transduction Domain

  • An, Songhie;Park, Jong-Sang
    • Bulletin of the Korean Chemical Society
    • /
    • v.34 no.9
    • /
    • pp.2589-2593
    • /
    • 2013
  • Gene therapy using nonviral gene delivery carriers has focused on the development and modification of synthetic carriers such as liposomes and polymers. Most polymers that are commercially used are taking advantage of their polycationic character which allows not only strong ligand-DNA affinity but also competent cell penetration. Despite the relatively high transfection efficiencies, high cytotoxicity is continuously pointed out as one of the major shortcomings of polycationic polymers such as PEI. Studies on the utilization of peptides have therefore been carried out recently to overcome these problems. For these reasons, the human transcription factor Hph-1, which is currently known as a protein transduction domain (PTD), was investigated in this study to evaluate its potential as a gene delivery carrier. Although its transfection efficiency was about 10-fold lower than PEI, it displayed almost no cytotoxicity even at concentrations as high as $100{\mu}M$. Hph-1 was oxidatively polymerized to yield poly-Hph-1. The cell viability of poly-Hph-1 transfected U87MG and NIH-3T3 cells was almost as high as the control (untreated) groups, and the transfection efficiency was about 10-fold higher than PEI. This study serves as a preliminary evaluation of Hph-1 and encourages further investigation.

The Heat Shock Protein 27 (Hsp27) Operates Predominantly by Blocking the Mitochondrial-Independent/Extrinsic Pathway of Cellular Apoptosis

  • Tan, Cheau Yih;Ban, Hongseok;Kim, Young-Hee;Lee, Sang-Kyung
    • Molecules and Cells
    • /
    • v.27 no.5
    • /
    • pp.533-538
    • /
    • 2009
  • Heat shock protein 27 (Hsp27) is a molecular chaperone protein which regulates cell apoptosis by interacting directly with the caspase activation components in the apoptotic pathways. With the assistance of the Tat protein transduction domain we directly delivered the Hsp27 into the myocardial cell line, H9c2 and demonstrate that this protein can reverse hypoxia-induced apoptosis of cells. In order to characterize the contribution of Hsp27 in blocking the two major apoptotic pathways operational within cells, we exposed H9c2 cells to staurosporine and cobalt chloride, agents that induce mitochondria-dependent (intrinsic) and -independent (extrinsic) pathways of apoptosis in cells respectively. The Tat-Hsp27 fusion protein showed a greater propensity to inhibit the effect induced by the cobalt chloride treatment. These data suggest that the Hsp27 predominantly exerts its protective effect by interfering with the components of the extrinsic pathway of apoptosis.

Enhancement of Adenoviral Transduction and Immunogenecity of Transgenes by Soluble Coxsackie and Adenovirus Receptor-TAT Fusion Protein on Dendritic Cells

  • Kim, Hye-Sung;Park, Mi-Young;Park, Jung-Sun;Kim, Chang-Hyun;Kim, Sung-Guh;Oh, Seong-Taek;Kim, Tai-Gyu
    • IMMUNE NETWORK
    • /
    • v.6 no.4
    • /
    • pp.192-198
    • /
    • 2006
  • Background: Investigating strategy to enhance efficiency of gene transfer via adenovirus is critical to sustain gene expression in targeted cells or tissues to regulate immune responses. However, the use of adenovirus as a gene delivery method has been limited by the native tropism of the virus. In this study, the critical parameter is to improve the efficient binding of viral particles to the plasma membrane prior to cellular uptake. Methods: Human immunodeficiency virus (HIV-1) trans-acting activator of transcription (TAT), a protein transduction domain, was fused to the ectodomain of the coxsackie-adenovirus receptor (CAR). The CAR-TAT protein was produced from a Drosophila Schneider 2 cells (S2) transfected with CAR-TAT genes. The function of CARTAT was analyzed the efficiency of adenoviral gene transfer by flow cytometry, and then immunizing AdVGFP with CAR-TAT was transduced on dendritic cells (DCs). Results: S2 transfectants secreting CAR-TAT fusion protein has been stable over a period of 6 months and its expression was verified by western blot. Addition of CAR-TAT induced higher transduction efficiency for AdVGFP at every MOI tested. When mice were vaccinated with DC of which adenoviral transduction was mediated by CAR-TAT, the number of IFN-${\gamma}$ secreting T-cells was increased as compared with those DCs transduced without CAR-TAT. Conclusion: Our data provide evidence that CAR-TAT fusion protein enhances adenoviral transduction and immunogenecity of transgenes on DCs and may influence on the development of adenoviral-mediated anti-tumor immunotherapy.

Update on Phosphorylation-Mediated Brassinosteroid Signaling Pathways (단백질 인산화에 의해 매개되는 브라시노스테로이드 신호전달 연구의 최근 상황)

  • Lee, Yew;Kim, Soo-Hwan
    • Journal of Life Science
    • /
    • v.22 no.3
    • /
    • pp.428-436
    • /
    • 2012
  • Protein phosphorylation is a universal mechanism that regulates cellular activities. The brassinosteroid (BR) signal transduction pathway is a relay of phosphorylation and dephosphorylation cascades. It starts with the BR-induced activation of the membrane receptor kinase brassinosteroid insensitive 1 (BRI1), resulting in the dephosphorylation of transcription factors such as BZR1/BES2 and BZR2/BES1 followed by BR-induced gene expression. Brassinosteroid signal transduction research has progressed rapidly by identifying the phosphorylation/dephosphorylation site(s) of the BR-regulated kinase and phosphatase substrates with a simultaneous pursuit of mutant phenotypes. Autophosphorylation, transphosphorylation, and serine/threonine and tyrosine phosphorylation of the receptor protein kinases BRI1 and BRI1-associated kinase (BAK1) have increased the understanding of the regulatory role of those kinases during physiological and developmental processes in plants. The phosphorylation event initiated by BR is also found in the regulation of receptor-mediated endocytosis and the subsequent degradation of the receptor. However, the basic molecular links of the BR signal transduction pathway are not well understood regarding this phosphorylation/dephosphorylation event. This review summarizes the current state of BR signal transduction research to uncover the phosphorylation/dephosphorylation networks and suggests directions for future research on steroid signal transduction to gain a more comprehensive understanding of the process.