• Title/Summary/Keyword: Functional molecules

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Integrated Quantitative Phosphoproteomics and Cell-Based Functional Screening Reveals Specific Pathological Cardiac Hypertrophy-Related Phosphorylation Sites

  • Kwon, Hye Kyeong;Choi, Hyunwoo;Park, Sung-Gyoo;Park, Woo Jin;Kim, Do Han;Park, Zee-Yong
    • Molecules and Cells
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    • v.44 no.7
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    • pp.500-516
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    • 2021
  • Cardiac hypertrophic signaling cascades resulting in heart failure diseases are mediated by protein phosphorylation. Recent developments in mass spectrometry-based phosphoproteomics have led to the identification of thousands of differentially phosphorylated proteins and their phosphorylation sites. However, functional studies of these differentially phosphorylated proteins have not been conducted in a large-scale or high-throughput manner due to a lack of methods capable of revealing the functional relevance of each phosphorylation site. In this study, an integrated approach combining quantitative phosphoproteomics and cell-based functional screening using phosphorylation competition peptides was developed. A pathological cardiac hypertrophy model, junctate-1 transgenic mice and control mice, were analyzed using label-free quantitative phosphoproteomics to identify differentially phosphorylated proteins and sites. A cell-based functional assay system measuring hypertrophic cell growth of neonatal rat ventricle cardiomyocytes (NRVMs) following phenylephrine treatment was applied, and changes in phosphorylation of individual differentially phosphorylated sites were induced by incorporation of phosphorylation competition peptides conjugated with cell-penetrating peptides. Cell-based functional screening against 18 selected phosphorylation sites identified three phosphorylation sites (Ser-98, Ser-179 of Ldb3, and Ser-1146 of palladin) displaying near-complete inhibition of cardiac hypertrophic growth of NRVMs. Changes in phosphorylation levels of Ser-98 and Ser-179 in Ldb3 were further confirmed in NRVMs and other pathological/physiological hypertrophy models, including transverse aortic constriction and swimming models, using site-specific phospho-antibodies. Our integrated approach can be used to identify functionally important phosphorylation sites among differentially phosphorylated sites, and unlike conventional approaches, it is easily applicable for large-scale and/or high-throughput analyses.

Downregulation of SETD5 Suppresses the Tumorigenicity of Hepatocellular Carcinoma Cells

  • Park, Mijin;Moon, Byul;Kim, Jong-Hwan;Park, Seung-Jin;Kim, Seon-Kyu;Park, Kihyun;Kim, Jaehoon;Kim, Seon-Young;Kim, Jeong-Hoon;Kim, Jung-Ae
    • Molecules and Cells
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    • v.45 no.8
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    • pp.550-563
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    • 2022
  • Hepatocellular carcinoma (HCC) is an aggressive and incurable cancer. Although understanding of the molecular pathogenesis of HCC has greatly advanced, therapeutic options for the disease remain limited. In this study, we demonstrated that SETD5 expression is positively associated with poor prognosis of HCC and that SETD5 depletion decreased HCC cell proliferation and invasion while inducing cell death. Transcriptome analysis revealed that SETD5 loss downregulated the interferon-mediated inflammatory response in HCC cells. In addition, SETD5 depletion downregulated the expression of a critical glycolysis gene, PKM (pyruvate kinase M1/2), and decreased glycolysis activity in HCC cells. Finally, SETD5 knockdown inhibited tumor growth in xenograft mouse models. These results collectively suggest that SETD5 is involved in the tumorigenic features of HCC cells and that targeting SETD5 may suppress HCC progression.

Human Vesicular Glutamate Transporters Functionally Complement EAT-4 in C. elegans

  • Lee, Dukgyu;Jung, Sunki;Ryu, Jungmin;Ahnn, Joohong;Ha, Ilho
    • Molecules and Cells
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    • v.25 no.1
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    • pp.50-54
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    • 2008
  • The vesicular glutamate transporter (VGLUT) transports glutamate into pre-synaptic vesicles. Three isoforms of VGLUT have been identified in humans, but their functional differences remain largely unknown. EAT-4 is the only homologue of human VGLUT in C. elegans. Here we report that mutants of eat-4 exhibit hyperforaging behavior and that each of the isoforms of human VGLUT functionally rescues the defects in eat-4 worms.

Structure-function analysis of PRDI DNA polymerase

  • Jung, Guhung
    • The Microorganisms and Industry
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    • v.16 no.3
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    • pp.6-14
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    • 1990
  • PRDI DNA polymerase is the smallest member of the family B DNA polymerase (Jung et al., 1987). This DNA polyerase is specified by bacteriophage PRDI which infects a wide variety of gram-negative bacteria(Mindich and Bamford, 1988). Because PRDI is highly amenable to genetic and biochemical manipulation, it is a convenient model system with which to study structure-function relationships of DNA polymerase molecules. To determine the functional roles of the highly conserved regions of the family B DNA polymerases, we have initiated site-directed mutagenesis with PRD1 DNA polymerase, and our results show that mutations at the conserved regions within PRD1 DNA polymerase inactivate polymerase complementing activity and catalytic activity.

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Electrical Properties of Organic Thin Films by Stimulus Speed (자극속도에 따른 유기박막의 전기 특성)

  • 박석순;김재민;정헌상;김창석;이권현;이경섭
    • Proceedings of the Korean Institute of Electrical and Electronic Material Engineers Conference
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    • 1997.04a
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    • pp.209-212
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    • 1997
  • Langmuir-Blodgett(LB) method is knowers as a unique method for preparing organic thin films, which can control thickness of the films in molecular level, and many kinds of ultra thin films of functional molecules have been prepared using this method. in this study, the electrical properties of phospholipid monolayers on a water surface was inve stigated by means of stimulus speed(40mm/min, 60mm/min).

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A Study on the Orientation Properties of Optical Opposite Organic Membrane (광이성 유기막의 배향특성에 관한 연구)

  • 박석순;조수영;전동규;이경섭;권영수
    • Proceedings of the Korean Institute of Electrical and Electronic Material Engineers Conference
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    • 1996.05a
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    • pp.248-251
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    • 1996
  • Langmuir-Biodgett(LB) method is known as a unique method for preparing organic thin films, which can control thickness of the films in molecular level, and many kinds of ultra thin films of functional molecules have been prepared using this method. In this study, The organization of phospholipid monolayers on a water surface was investigated by means of displacement current measurement technique.

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Effect of hydrogen on adsorption of hydrocarbon fragments on graphene

  • Cho, Sangmo
    • Proceeding of EDISON Challenge
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    • 2014.03a
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    • pp.464-466
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    • 2014
  • We investigate the effect of hydrogen on adsorption of hydrocarbon molecules on graphene with density functional theory (DFT) calculations. In this study, we calculate the binding energies of hydrogen molecule, carbon atom and other hydrocarbon fragments such as CHx (x=1, 2, 3, 4) on graphene to find the most stable adsorption site. Then, to study the effect of hydrogen, we investigate the adsorption of hydrocarbon fragments in the presence of hydrogen atoms on graphene.

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Virtual Screening, Docking and DFT Study of PRMT5

  • Subathra, S
    • Journal of Integrative Natural Science
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    • v.15 no.4
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    • pp.187-194
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    • 2022
  • Protein Arginine Methyltransferase 5 (PRMT5), a significant member of the PRMT family, is a promising anticancer target. In this study, novel small compounds that act against the PRMT5 target are found by combining virtual screening with ChEMBL database medicines and Density Functional Theory. The ChEMBL database compounds were screened to retrieve the hit molecules, which further subjected for DFT analysis. Finally we have evaluated that ChEMBL- approved drugs such as Lifitegrast, Abiraterone acetate and Solifenacin may be potential inhibitors for PRMT5.

Aptamers as Functional Nucleic Acids: in vitro Selection and Biotechnological Applications

  • You, Kyung-Man;Lee, Sang-Hyun;Aesul Im;Lee, Sun-Bok
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.8 no.2
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    • pp.64-75
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    • 2003
  • Aptamers are functional nucleic acids that can specially bind to proteins, peptides, amino acids. nucleotides, drugs, vitamins and other organic and inorganic compounds. The aptamers are identified from random DNA or RNA libraries by a SELEX (systematic evolution of ligands by exponential amplification) process. As aptamers have the advantage, and potential ability to be released from the limitations of antibodies, they are attractive to a wide range of therapeutic and diagnostic applications. Aptamers, with a high-affinity and specificity, could fulfil molecular the recognition needs of various fields in biotechnology. In this work, we reviewed some aptamer Selection techniques, properties, medical applications of their molecules and their biotechnological applications, such as ELONA (enzyme linked oligonucleotide assay), flow cytometry, biosensors, electrophoresis, chromatography and microarrays.