• Title/Summary/Keyword: 단백질체

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Identification of Diseasomal Proteins from Atopy-Related Disease Network (아토피관련 질병 네트워크로부터 질병단백체 발굴)

  • Lee, Yoon-Kyeong;Yeo, Myeong-Ho;Kang, Tae-Ho;Yoo, Jae-Soo;Kim, Hak-Yong
    • The Journal of the Korea Contents Association
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    • v.9 no.4
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    • pp.114-120
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    • 2009
  • In this study, we employed the idea that disease-related proteins tend to be work as an important factor for architecture of the disease network. We initially obtained 43 atopy-related proteins from the Online Mendelian Inheritance in Man (OMIM) and then constructed atopy-related protein interaction network. The protein network can be derived the map of the relationship between different disease proteins, denoted disease interaction network. We demonstrate that the associations between diseases are directly correlated to their underlying protein-protein interaction networks. From constructed the disease-protein bipartite network, we derived three diseasomal proteins, CCR5, CCL11, and IL/4R. Although we use the relatively small subnetwork, an atopy-related disease network, it is sufficient that the discovery of protein interaction networks assigned by diseases will provide insight into the underlying molecular mechanisms and biological processes in complex human disease system.

Overproduction and Purification of Ribose-Binding Proteins from the Wild-Type Mutant and Revertant Strains in Escherichia coli (리보스 결합단백질의 대량생산을 위한 야생형 수송결합변이, 복귀변이 유전자의 클로닝과 이들 단백질의 순수정제)

  • ;Randall Linda L.
    • Korean Journal of Microbiology
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    • v.26 no.4
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    • pp.291-297
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    • 1988
  • Three alleles of rbsB gene, rbsB, rbsB103, and rbsB106 from the wild type, the mutant and the revertant strain, respectively, were cloned for overproduction of proteins under the control of lambda $P_{L}$ promoter. Five different species of precursor and mature ribose-binding proteins were purified to homogeneity using DEAE-Sephadex column chromatography, osmotic shock pocedure, CM-Sephadex column chromatography, and Chromatofocusing column chromaography. pI of the precursor proteins and mature proteins were determined and found to be pH 8.0 and 7.5, respectively. The purified proteins were subjected to amino acid sequencing. The results confirmed the amino acid changes deduced from the DNA sequencing.

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Construction of a Transgenic Plant to Develop a New Method for the Isolation of Calmodulin-Binding Proteins (새로운 방법을 이용한 칼모둘린 결합 단백질 분리를 위한 형질 전환 식물체의 구축)

  • Kim, Sun-Ho;Lee, Kyung-Hee;Kim, Kyung-Eun;Jung, Mi-Soon;Lim, Chae-Oh;Lee, Shin-Woo;Chung, Woo-Sik
    • Journal of Life Science
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    • v.17 no.9 s.89
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    • pp.1177-1181
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    • 2007
  • Calmodulin (CaM), a ubiquitous calcium-binding protein, regulates diverse cellular functions by modulating the activity of a variety CaM-binding proteins (CaMBPs). Because eukaryotes have multiple CaMBPs, it is important to isolate and characterize them in different tissues and conditions. So far a number of CaMBPs have been identified through classical screening methods. Many classes of proteins have been predicted to bind CaMs based on their structural homology with already known targets. In an effort to develop a method for large-scale analysis of CaMBPs in Arabidopsis, we have generated a transgenic plants overexpressing AtCaM2-GFP. We performed protein pull-down assay to test whether exogenously expressed AtCaM2-GFP proteins can interact with CaMBPs. The exogenously expressed AtCaM2-GFP could strongly interact with a CaMBP, AS1 protein. This result suggests that AtCaM2-GFP in transgenic plants may interact with many CaMBPs in plant cell. Therefore, we will be able to isolate kinds of CaMBPs by using these transgenic plants in many different tissue and environments.

The Scaffolding Protein WAVE1 Associates with Kinesin 1 through the Tetratricopeptide Repeat (TPR) Domain of the Kinesin Light Chain (KLC) (Kinesin Light Chain (KLC)의 Tetratricopeptide Repeat (TPR) 도메인을 통한 Scaffold 단백질 WAVE1과 Kinesin 1의 결합)

  • Jang, Won Hee;Jeong, Young Joo;Urm, Sang-Hwa;Seog, Dae-Hyun
    • Journal of Life Science
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    • v.26 no.8
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    • pp.963-969
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    • 2016
  • Kinesin superfamily proteins (KIFs) are microtubule-dependent molecular motor proteins essential for the intracellular transport of organelles and protein complexes in cells. Kinesin 1 is a member of those KIFs that transport various cargoes, including organelles, synaptic vesicles, neurotransmitter receptors, cell signaling molecules, and mRNAs through interaction between its light chain subunit and the cargoes. Kinesin light chains (KLCs) are non-motor subunits that associate with the kinesin heavy chain (KHC) dimer. KLCs interact with many different binding proteins, but their particular binding proteins have not yet been fully identified. We used the yeast two-hybrid assay to identify proteins that interact with the tetratricopeptide repeat (TPR) domain of KLC1. We found an interaction between the TPR domain of KLC1 and Wiskott-Aldrich syndrome protein family member 1 (WAVE1), a member of the WASP/WAVE family involved in regulation of actin cytoskeleton. WAVE1 bound to the six TPR domain-containing regions of KLC1 and did not interact with KHCs (KIF5A, KIF5B, and KIF5C) in the yeast two-hybrid assay. The carboxyl (C)-terminal verprolin-cofilin-acidic (VCA) domain of WAVE1 is essential for interaction with KLC1. Also, other WAVE isoforms (WAVE2 and WAVE3) interacted with KLC1 in the yeast two-hybrid assay. When co-expressed in HEK-293T cells, WAVE1 co-localized with KLC1 and co-immunoprecipitated with KLC1 and KIF5B. These results suggest that kinesin 1 motor protein may transport WAVE complexes or WAVE-coated cargoes in cells.

RGS3 Suppresses cAMP Response Element (CRE) Activity Mediated by CB2 Cannabinoid Receptor in HEK293 Cells (캐너비노이드 수용체 CB2의 신호전달작용에 미치는 RGS3의 억제적 효과)

  • Kim, Sung-Dae;Lee, Whi-Min;Endale, Mehari;Cho, Jae-Youl;Park, Hwa-Jin;Oh, Jae-Wook;Rhee, Man-Hee
    • Journal of Life Science
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    • v.19 no.11
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    • pp.1506-1513
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    • 2009
  • RGS proteins have been identified as negative regulators of G protein signalling pathways and attenuate the activity of GPCR receptors. However, information on the regulatory effects of RGS proteins in the activity of cannabinoid receptors is limited. In this study, the role of RGS proteins on the signal transduction of the CB2 cannabinoid receptor was investigated in HEK293 cells co-transfected with CB2-receptors and plasmids encoding RGS2, RGS3, RGS4 and RGS5. Treatment of cells with WIN55, 212-2, a CB2 receptor agonist, inhibited forskolin-induced cAMP response element (CRE) activity in CB2-transfected HEK293 (CB2-HEK293) cells. This inhibitory effect of WIN 55, 212-2 on CRE activity was reversed by co-transfection of CB2-HEK293 cells with RGS3, but not with RGS2, RGS4 and RGS5. However, endogenous RGS3 protein knocked down by a small interfering siRNA targeting RGS3 gene enhanced inhibition of forskolin induced CRE activity via agonist induced CB2 receptor signal transduction. These results indicate the functional role of endogenous RGS protein in cannabinoid signaling pathways and define receptor-selective roles of endogenous RGS3 in modulating CRE transcriptional responses to agonist induced CB2 receptor activity.

Gene Expression in The Fifth Generation of TMV Resistant Transgenic Tobacco Plane at Elevated Temperature (TMV 저항성 형질전환 연초식물체 제 5 세대에서 유전자 안정성 및 고온조건에서의 유전자 발현)

  • 이기원;박성원;이청호;박은경;김상석;최순용
    • Korean Journal of Plant Tissue Culture
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    • v.25 no.4
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    • pp.245-250
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    • 1998
  • Tobacco mosaic virus(TMV) coat protein cDNA was transformed to Nicotiana tabacum cv. NC82 and the transgenic tobacco plants resistant to TMV infection were isolated in the next generation. The expression of TMV coat protein cDNA and genetic stability of the fifth generation of TMV resistant transgenic tobacco plants at the higher temperature were investigated. The TMV coat protein cDNA was amplified by genomic PCR in all the TMV resistant transgenic tobacco plants. The TMV coat protein expressed in the transgenic tobacco plants was detected at very low level by immunoblot hybridization. Even in tansgenic plants that showed the viral symptom only on very late sucker growth (delay type plants), the coat protein expression in the suckers was much less than that of susceptible tobacco infected with TMV. The TMV coat protein expressed in the transgenic tobacco plants was below 0.01% of total protein. Transcription and expression of the coat protein cDNA in delay type plants were observbed at high temperature (38$^{\circ}C$), and TMV replication was suppressed at both 28$^{\circ}C$ and 38$^{\circ}C$. This indicates that unlike the resistance conferred by 'N' gene. TMV resistance of transgenic tobacco plant won't break down at high temperature.

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Development of Hydrogel for Decrease Protein Adsorption and Application to Intraocular Lens (단백질흡착 감소용 하이드로겔의 개발 및 인공수정체로의 적용)

  • Ko, Na Young;Lee, Hyun Mee
    • Journal of the Korean Chemical Society
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    • v.62 no.1
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    • pp.30-35
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    • 2018
  • The purpose of this study was to decrease the protein adsorption and improve the function of the hydrophobic acrylic Intraocular lens(IOL). Hydrophobic acrylic intraocular lenses were prepared by using ethyleneglycol phenyletheracrylate (EGPEA), styrene and 2-hydroxyethyl methacrylate (HEMA). Polyvinyl pyrrolidone (PVP) and 2-methacryloyloxyethyl phosphorylcholine (MPC) were used as additives. Water contents, wettability, light transmittance and protein adsorption amount were measured to evaluate the physical properties of the intraocular lens. The water content and wettability of all samples containing additives were increased and the amount of protein adsorption decreased. In particular, samples containing MPC showed a further decrease in protein adsorption. The hydrophobic acrylic intraocular lens with PVP and MPC was found to improve the function of the intraocular lens by reducing the protein adsorption while having basic physical properties.

Immunocytochemical Investigation on the Intracisternal Accumulations of Storage Protein in Pea Cotyledon Cells (완두 자엽세포의 소포체 내강에 축적된 저장 단백질에 대한 면역세포화학적 연구)

  • Jeong, Byung-Kap;Park, Hong-Duok
    • Applied Microscopy
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    • v.31 no.2
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    • pp.199-206
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    • 2001
  • In 1980s, the fragmentation or subdivision of protein deposits at the periphery of protein storage vacuole was suggested as the only route of PB development in pea cotyledon cells. Since then, other independant processes such as terminal dilation , transformation and de novo development have been discussed as alternative routes for PB development, and today, these multiple mechanisms of PB development are accepted as a result of active investigations. For analysis of the protein accumulations in the ER cisternae during seed development, immunocytochemical gold labellings were applyed on the single cells separated by enzymatic digestion from cotyledon tissue. Anti-legumin labellings at the early stage, and anti-vicilin labellings at the intermediate stage were observed on the protein-filled ER. The $\alpha-Tip$, which is the ER retention protein, was labelled somewhat at late stage, and PPase, a sort of tonoplast membrane protein, was labelled at early stage.

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