• Title/Summary/Keyword: trafficking

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A Machine Learning Based Method for the Prediction of G Protein-Coupled Receptor-Binding PDZ Domain Proteins

  • Eo, Hae-Seok;Kim, Sungmin;Koo, Hyeyoung;Kim, Won
    • Molecules and Cells
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    • v.27 no.6
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    • pp.629-634
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    • 2009
  • G protein-coupled receptors (GPCRs) are part of multi-protein networks called 'receptosomes'. These GPCR interacting proteins (GIPs) in the receptosomes control the targeting, trafficking and signaling of GPCRs. PDZ domain proteins constitute the largest protein family among the GIPs, and the predominant function of the PDZ domain proteins is to assemble signaling pathway components into close proximity by recognition of the last four C-terminal amino acids of GPCRs. We present here a machine learning based approach for the identification of GPCR-binding PDZ domain proteins. In order to characterize the network of interactions between amino acid residues that contribute to the stability of the PDZ domain-ligand complex and to encode the complex into a feature vector, amino acid contact matrices and physicochemical distance matrix were constructed and adopted. This novel machine learning based method displayed high performance for the identification of PDZ domain-ligand interactions and allowed the identification of novel GPCR-PDZ domain protein interactions.

Biochemistry, Molecular Biology, and Metabolic Engineering of Benzylisoquinoline Alkaloid Biosynthesis

  • Peter J. Facchini;Park, Sang-Un;David A. Bird;Nailish Samanani
    • Korean Journal of Plant Tissue Culture
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    • v.27 no.4
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    • pp.269-282
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    • 2000
  • Benzylisoquinoline alkaloids are a diverse group of natural products that include many pharmacologically active compounds produced in a limited number of plant families. Despite their complexity, intensive biochemical research has extended our knowledge of the chemistry and enzymology of many important benzylisoquinoline alkaloid pathways, such as those leading to the analgesic drugs morphine and codeine, and the antibiotics sanguinarine and berberine. The use of cultured plant cells as an experimental system has facilitated the identification and characterization of more than 30 benzylisoquinoline alkaloid biosynthetic enzymes, and the molecular cloning of the genes that encode at least 8 of these enzymes. The recent expansion of biochemical and molecular technologies has creat-ed unique opportunities to dissect the mechanisms involved in the regulation of benzylisoquinoline alkaloid biosynthesis in plants. Research has suggested that product accumulation is controlled by the developmental and inducible regulation of several benzylisoquinoline alkaloid biosynthetic genes, and by the subcellular compartmentation of biosynthetic enzymes and the intracellular localization and trafficking of pathway intermediates. In this paper, we review our current understanding of the biochemistry, cell biology, and molecular regulation of benzylisoquinoline alkaloid biosynthesis in plants. We also summarize our own research activities, especially those related to the establishment of protocols for the genetic transformation of benzylisoquinoline alkaloid-producing species, and the development of metabolic engineering strategies in these plants.

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Affinity between TBC1D4 (AS160) phosphotyrosine-binding domain and insulin-regulated aminopeptidase cytoplasmic domain measured by isothermal titration calorimetry

  • Park, Sang-Youn;Kim, Keon-Young;Kim, Sun-Min;Yu, Young-Seok
    • BMB Reports
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    • v.45 no.6
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    • pp.360-364
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    • 2012
  • Uptake of circulating glucose into the cells happens via the insulin-mediated signalling pathway, which translocates the glucose transporter 4 (GLUT4) vesicles from the intracellular compartment to the plasma membrane. Rab GTPases are involved in this vesicle trafficking, where Rab GTPases-activating proteins (RabGAP) enhance the GTP to GDP hydrolysis. TBC1D4 (AS160) and TBC1D1 are functional RabGAPs in the adipocytes and the skeletonal myocytes, respectively. These proteins contain two phosphotyrosine-binding domains (PTBs) at the amino-terminus of the catalytic RabGAP domain. The second PTB has been shown to interact with the cytoplasmic region of the insulin-regulated aminopeptidase (IRAP) of the GLUT4 vesicle. In this study, we quantitatively measured the ${\sim}{\mu}M$ affinity ($K_D$) between TBC1D4 PTB and IRAP using isothermal titration calorimetry, and further showed that IRAP residues 1-49 are the major region mediating this interaction. We also demonstrated that the IRAP residues 1-15 are necessary but not sufficient for the PTB interaction.

Host Cell Nuclear Localization of Shigella flexneri Effector OspF Is Facilitated by SUMOylation

  • Jo, Kyungmin;Kim, Eun Jin;Yu, Hyun Jin;Yun, Cheol-Heui;Kim, Dong Wook
    • Journal of Microbiology and Biotechnology
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    • v.27 no.3
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    • pp.610-615
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    • 2017
  • When Shigella infect host cells, various effecter molecules are delivered into the cytoplasm of the host cell through the type III secretion system (TTSS) to facilitate their invasion process and control the host immune responses. Among these effectors, the S. flexneri effector OspF dephosphorylates mitogen-activated protein kinases and translocates itself to the nucleus, thus preventing histone H3 modification to regulate expression of proinflammatory cytokines. Despite the critical role of OspF, the mechanism by which it localizes in the nucleus has remained to be elucidated. In the present study, we identified a potential small ubiquitin-related modifier (SUMO) modification site within OspF and we demonstrated that Shigella TTSS effector OspF is conjugated with SUMO in the host cell and this modification mediates the nuclear translocation of OspF. Our results show a bacterial virulence factor can exploit host post-translational machinery to execute its intracellular trafficking.

Priority Analysis of Sub-goals Related to Infants and Young Children in Korean Sustainable Development Goals (K-SDGs) Using the Delphi Technique (델파이 기법을 활용한 국가지속가능발전목표(K-SDGs)의 영유아 관련 세부목표의 우선순위 분석)

  • Jeon, Hyo Jeong;Goh, Eun Kyoung
    • Korean Journal of Childcare and Education
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    • v.16 no.3
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    • pp.1-17
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    • 2020
  • Objective: The purpose of this study is to identify and prioritize sub-goals for infants and young children among the Korean Sustainable Development Goals (K-SDGs). Methods: The expert panel of this study consisted of 15 faculty members from the department of early childhood education or children's studies at 12 universities and was put together in order to conduct a Delphi survey. Results: As a result of the study, first, a total of 16 sub-goals were developed in order to target infants and young children. Second, the 16 sub-goals were then ranked according to their importance and urgency, and the most important goal were 'the improvement of mental health and prevention of drug abuse (K-SDG 3-2)'. And 'the elimination of violence and discrimination against girls (K-SDG 5-1, 5-2)', 'the safe and inclusive learning environment (K-SDG 4-8)' and 'end of abuse, trafficking, exploitation and all forms of violence against children (K-SDG 16-2)' were the next important and urgent goals. These priority objectives reflect the inviolable rights that infants and young children in Korea should be guaranteed by default. Conclusion/Implications: This study provided a basis for its effective implementation by identifying goals and priorities for infants and young children in comprehensive and interdependent K-SDGs.

PKD2 interacts with Lck and regulates NFAT activity in T cells

  • Li, Qing;Sun, Xiaoqing;Wu, Jun;Lin, Zhixin;Luo, Ying
    • BMB Reports
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    • v.42 no.1
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    • pp.35-40
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    • 2009
  • Protein kinase D2 (PKD2) is a member of the PKD serine/threonine protein kinase family that has been implicated in the regulation of a variety of cellular processes including proliferation, survival, protein trafficking and immune response. In the present study, we report a novel interaction between PKD2 and Lck, a member of the Src tyrosine protein kinase family that is predominantly expressed in T cells. This interaction involved the C-terminal kinase domains of both PKD2 and Lck. Moreover, co-expression of Lck enhanced the tyrosine phosphorylation of PKD2 and increased its kinase activity. Finally, we report that PKD2 enhanced T cell receptor (TCR)-induced nuclear factor of T cell (NFAT) activity in Jurkat T cells. These results suggested that Lck regulated the activity of PKD2 by tyrosine phosphorylation, which in turn may have modulated the physiological functions of PKD2 during TCR-induced T cell activation.

The Plant Cellular Systems for Plant Virus Movement

  • Hong, Jin-Sung;Ju, Ho-Jong
    • The Plant Pathology Journal
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    • v.33 no.3
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    • pp.213-228
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    • 2017
  • Plasmodesmata (PDs) are specialized intercellular channels that facilitate the exchange of various molecules, including sugars, ribonucleoprotein complexes, transcription factors, and mRNA. Their diameters, estimated to be 2.5 nm in the neck region, are too small to transfer viruses or viral genomes. Tobacco mosaic virus and Potexviruses are the most extensively studied viruses. In viruses, the movement protein (MP) is responsible for the PD gating that allows the intercellular movement of viral genomes. Various host factors interact with MP to regulate complicated mechanisms related to PD gating. Virus replication and assembly occur in viral replication complex (VRC) with membrane association, especially in the endoplasmic reticulum. VRC have a highly organized structure and are highly regulated by interactions among the various host factors, proteins encoded by the viral genome, and the viral genome. Virus trafficking requires host machineries, such as the cytoskeleton and the secretory systems. MP facilitates the virus replication and movement process. Despite the current level of understanding of virus movement, there are still many unknown and complex interactions between virus replication and virus movement. While numerous studies have been conducted to understand plant viruses with regards to cell-to-cell movement and replication, there are still many knowledge gaps. To study these interactions, adequate research tools must be used such as molecular, and biochemical techniques. Without such tools, virologists will not be able to gain an accurate or detailed understanding of the virus infection process.

Recent advances in intravital microscopy for investigation of dynamic cellular behavior in vivo

  • Choo, Yeon Woong;Jeong, Juhee;Jung, Keehoon
    • BMB Reports
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    • v.53 no.7
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    • pp.357-366
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    • 2020
  • Currently, most biological research relies on conventional experimental techniques that allow only static analyses at certain time points in vitro or ex vivo. However, if one could visualize cellular dynamics in living organisms, that would provide a unique opportunity to study key biological phenomena in vivo. Intravital microscopy (IVM) encompasses diverse optical systems for direct viewing of objects, including biological structures and individual cells in live animals. With the current development of devices and techniques, IVM addresses important questions in various fields of biological and biomedical sciences. In this mini-review, we provide a general introduction to IVM and examples of recent applications in the field of immunology, oncology, and vascular biology. We also introduce an advanced type of IVM, dubbed real-time IVM, equipped with video-rate resonant scanning. Since the realt-ime IVM can render cellular dynamics with high temporal resolution in vivo, it allows visualization and analysis of rapid biological processes.

Molecular Genetics and Diagnostic Approach of Mucolipidosis II/III

  • Sohn, Young Bae
    • Journal of mucopolysaccharidosis and rare diseases
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    • v.2 no.1
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    • pp.13-16
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    • 2016
  • Mucolipidosis (ML) II/III are autosomal recessive diseases caused by deficiency of post-translational modification of lysosomal enzymes. The mannose-6-phosphate (M6P) residue in lysosomal enzymes synthesized by N-acetylglucosamine 1-phosphotransferase (GlcNAc-phosphotransferase) serves as recognition marker for trafficking in lysosomes. GlcNAc-phosphotransferase is encoded by GNPTAB and GNPTG. Mutations in GNPTAB cause severe ML II alpha/beta and the attenuated ML III alpha/beta. Whereas mutations in GNPTG cause the ML III gamma, the attenuated type of ML III variant. For the diagnostic approaches, increased urinary oligosaccharides excretion could be a screening test in clinically suspicious patients. To confirm the diagnosis, instead of measuring the activity of GlcNAc phosphotransferase, measuring the enzymatic activities of different lysosomal hydrolases are useful for diagnosis. The activities of several lysosomal hydrolases are decreased in fibroblasts but increased in serum of the patients. In addition, the sequence analysis of causative gene is warranted. Therefore, the confirmatory diagnosis requires a combination of clinical evaluation, biochemical and molecular genetic testing. ML II/III show complex disease manifestations with lysosomal storage as the prime cellular defect that initiates consequential organic dysfunctions. As there are no specific therapy for ML to date, understanding the molecular pathogenesis can contribute to develop new therapeutic approaches ultimately.

In vivo Dendritic Cell Migration Tracking Using Near-infrared (NIR) Imaging (Near-infrared (NIR) 영상기법을 이용한 생체 내 수지상세포의 이동)

  • Lee, Jun-Ho;Jung, Nam-Chul;Lee, Eun Gae;Lim, Dae-Seog
    • KSBB Journal
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    • v.27 no.5
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    • pp.295-300
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    • 2012
  • Matured dendritic cells (DCs) begin migration with their release from the bone marrow (BM) into the blood and subsequent traffic into peripheral lymphoid and non-lymphoid tissues. Throughout this long movement, migrating DCs must apply specialized skills to reach their target destination. Non-invasive in vivo cell-tracking techniques are necessary to advance immune cell-based therapies. In this study, we used a DiD cell-tracking solution for in vivo dendritic cell tracking in naive mice. We tracked DiD (non-invasive fluorescence dye)-labeled mature dendritic cells using the Near Infrared (NIR) imaging system in normal mice. We examined the immunophenotype of DiD-labeled cells compared with non-labelled mature DCs, and obtained time-serial images of NIR-DC trafficking after mouse footpad injection. In conclusion, we confirmed that DiD-labeled DCs migrated into the popliteal lymph node 24 h after the footpad injection. Here, these data suggested that the cell tracking system with the stable fluorescence dye DiD was useful as a cell tracking tool to advance dendritic cell-based immunotherapy.