• Title/Summary/Keyword: Hybrid protein

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Identification of a Domain in Yeast Chitin Synthase 3 Interacting with Chitin Synthase 4 by Two-Hybrid Analysis

  • Park, Hyun-Sook;Shin-Jung-Choi;Nok-Hyun-Park;Chi-Hwa-Kim;Sung-Uk-Kim
    • Journal of Microbiology and Biotechnology
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    • v.12 no.6
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    • pp.943-949
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    • 2002
  • It has been proposed that chitin synthase 3 (CHS3)-nediated chitin synthesis during the vegetative cell cycle is regulated by chitin synthase 4 (CHS4) of Saccharomyces cerevisiae. To investigate direct protein-protein interaction between the coding products of these two genes, a domain of Chs3p that is responsible for interaction with Chs4p was identified, using the yeast two-hybrid system. This domain of 54 amino acids, termed MIRC3-4 (Maximum Interacting Region of Chs3p with Chs4p), is well conserved among CHS3 homologs of various fungi. Some mutations in MIRC3-4 resulted in a decrease in the enzymatic activity and chitin contents. Chs3p carrying those mutations exhibited weak interactions with Chs4p, when assayed by the yeast two-hybrid system. Surprisingly, all the mutants were sensitive to Calcofluor regardless of changes in enzymatic activities or chitin contents. This report deals with a core region in MIRC3-4 that affects the interaction with Chs4p.

An Analysis System for Protein-Protein Interaction Data Based on Graph Theory (그래프 이론 기반의 단백질-단백질 상호작용 데이타 분석을 위한 시스템)

  • Jin Hee-Jeong;Yoon Ji-Hyun;Cho Hwan-Gue
    • Journal of KIISE:Computer Systems and Theory
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    • v.33 no.5
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    • pp.267-281
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    • 2006
  • PPI(Protein-Protein Interaction) data has information about the organism has maintained a life with some kind of mechanism. So, it is used in study about cure research back, cause of disease, and new medicine development. This PPI data has been increased by geometric progression because high throughput methods are developed such as Yeast-two-hybrid, Mass spectrometry, and Correlated mRNA expression. So, it is impossible that a person directly manage and analyze PPI data. Fortunately, PPI data is able to abstract the graph which has proteins as nodes, interactions as edges. Consequently, Graph theory plentifully researched from the computer science until now is able to be applied to PPI data successfully. In this paper, we introduce Proteinca(PROTEin INteraction CAbaret) workbench system for easily managing, analyzing and visualizing PPI data. Proteinca assists the user understand PPI data intuitively as visualizing a PPI data in graph and provide various analytical function on graph theory. And Protenica provides a simplified visualization with gravity-rule.

Glutamate Receptor-interacting Protein 1 Protein Binds to the Armadillo Family Protein p0071/plakophilin-4 in Brain (Glutamate receptor-interacting protein 1 단백질과 armadillo family 단백질 p0071/plakophilin-4와의 결합)

  • Moon, Il-Soo;Seog, Dae-Hyun
    • Journal of Life Science
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    • v.19 no.8
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    • pp.1055-1061
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    • 2009
  • ${\alpha}$-amino-3-hydroxy-5-methyl-4-isoxazole propionate (AMPA) receptors are widespread throughout the central nervous system and appear to serve as synaptic receptors for fast excitatory synaptic transmission mediated by glutamate. Their modulation is believed to affect learning and memory. To identify the interaction proteins for the AMPA receptor subunit glutamate receptor-interacting protein 1 (GRIPl), GRIP1 interactions with armadillo family protein p0071/plakophilin-4 were investigated. GRIP1 protein bound to the tail region of p0071/plakophilin-4 but not to other armadillo family protein members in a yeast two-hybrid assay. The "S-X-V" motif at the carboxyl (C)-terminal end of p0071/plakophilin-4 is essential for interaction with GRIP1. p0071/plakophilin-4 interacted with the Postsynaptic density-95/Discs large/Zona occludens-1 (PDZ) domains of GRIPI in the yeast two-hybrid assay, as is indicated also by Glutathione S-transferase (GST) pull-down, and co-immunoprecipitated with GRIP1 antibody in brain fraction. The findings of this study provide evidence that p0071/plakophilin-4 is an interactor of GRIP1.

A Comparative Study on Nutritional Composition of Native and Hybrid Pork in Korea (재래종과 개량종 돼지고기의 영양성분 비교 연구)

  • 한재숙;한경필;소궤신행;여정수;이승언;일본명
    • Journal of the East Asian Society of Dietary Life
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    • v.13 no.3
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    • pp.185-190
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    • 2003
  • The purpose of this study was to investigate the nutritional composition of native and hybrid pork in Korea. Three different parts (ham, loin and belly) of both native and hybrid pork were used. The results were as follows The moisture content from Korean native pork was about 60.28%, while that from the loin of hybrid one was 69% and decreased in the order of him, loin, and belly The highest protein content of 19.71% was found in Korean native pork loin, and Korean native pork ham had a significant amount of protein of 17.80% and hybrid one had 13.14% (p< .05). The highest crude lipid, 34.44%, was found in hybrid pork belly, Korean native pork ham had a significant amount of 5.43% and hybrid pork had 2.33% (p< .05). The highest carbohydrate content of 13.28% was found in the Korean native pork belly. The amount of ash was in the order of loin, ham and belly in Korean native pork. Among the minerals, K was found the most in Korean native pork ham (654.82mg) and hybrid one (747. 35mg) (p< .05). Fe was higher in the Korean native pork ham (23.03mg), loin (15.86mg) and belly (10.80mg) compared to the hybrid pork ham (19.04mg), loin (11.63mg) and belly (7.61mg). That was significant ham, loin(p< .01) and belly(p< .05). The main free amino acids of the native and the hybrid pork in Korea were alanine, aspartic acid and lysine. While the cholesterol content was found to be high in the order of ham, belly, and loin in the Korean native pork, in the order of belly, fresh ham and loin in the hybrid pork. The cholesterol contents in ham were significantly different between the Korean native pork(789.32$\mu\textrm{g}$) and the hybrid pork (538.84$\mu\textrm{g}$) (p< .01).

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Direct Interaction of KIF5s and Actin-Based Transport Motor, Myo9s (KIF5s와 직접 결합하는 액틴 결합 운동단백질 Myo9s의 규명)

  • Seog, Dae-Hyun
    • Journal of Life Science
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    • v.21 no.8
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    • pp.1076-1082
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    • 2011
  • Microtubule-based kinesin motor proteins are used for long-range vesicular transport. KIF5s (KIF5A, KIF5B and KIF5C) mediate the transport of various membranous vesicles along microtubules, but the mechanism behind how they recognize and bind to a specific cargo has not yet been completely elucidated. To identify the interaction protein for KIF5B, yeast two-hybrid screening was performed and a specific interaction with the unconventional myosin Myo9b, an actin-based vesicle transport motor, was found. The GTPase-activating protein (GAP) domain of Myo9s was essential for interaction with KIF5B in the yeast two-hybrid assay. Myo9b bound to the carboxyl-terminal region of KIF5B and to other KIF5 members. In addition, glutathione S-transferase (GST) pull-downs showed that Myo9s specifically interact to the complete Kinesin-I complex. An antibody to KIF5B specifically co-immunoprecipitated KIF5B associated with Myo9s from mouse brain extracts. These results suggest that kinesin-I motor protein interacts directly with actin-based motor proteins in the cell.

Prediction of the Concentration of Diphenylhydantion in the Brain Using a Physiological Pharmacokinetic Hybrid Model

  • Song, Sae-Heum;Shim, Chang-Koo;Lee, Min-Hwa;Kim, Shin-Keun
    • Archives of Pharmacal Research
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    • v.13 no.3
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    • pp.221-226
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    • 1990
  • A physiological pharmacokinetic hybrid model was developed in order to predict the disposition kinetics of diphenylhydantoin (DPH) in the brain from the plasma conentration data of DPH. The model was constructed under the assumptions of well-stirred, plasma flow-limited and lienar tissue diposition kinetics of DPH. DPH was administered intravenously to the rats at a dose of 10 mg/kg together with/without sodium salicylate (SA;10 mg/kg) and the DPH concentrations in the plasma and brain were determined. Plasma protein binding of DPH concentrations in the plasma and brain were determined. Plasma protein binding of DPH was also determined using equilibrium dialysis technique. Then the model was tested for its predictability of DPH concentrations in the brian from the plasma data of DPH. It was found that the predicted values of DPH concentrations in the brian were in fair agreement with the experimental values in the rats of both treatments. The 2-fold increase in the brain concentration of DPH by SA-coadinistration was predicted well from the plasma concentration and plasma free fraction ($f_p$) data of DPH using the model. Therefore, the hybrid model was concluded to be very useful for the prediction of the concentrations of DPH in the brain from the plasma concentration data. Finally, DPH concentrations in the human brian was calculated using this model from plasma DPH data in the literature, yet the scale-up of this model to the human is not convinced.

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Homo- or Hetero-Dimerization of Muscarinic Receptor Subtypes is Not Mediated by Direct Protein-Protein Interaction Through Intracellular and Extracellular Regions

  • Kang, Yun-Kyung;Yoon, Tae-Sook;Lee, Kyung-Lim;Kim, Hwa-Jung
    • Archives of Pharmacal Research
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    • v.26 no.10
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    • pp.846-854
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    • 2003
  • The oligomerization of G-proteincoupled receptors (GPCRs) has been shown to occur by various mechanisms, such as via disulfide covalent linkages, non covalent (ionic, hydrophobic) interactions of the N-terminal, and/or transmembrane and/or intracellular domains. Interactions between GPCRs could involve an association between identical proteins (homomers) or non-identical proteins (heteromers), or between two monomers (to form dimers) or multiple monomers (to form oligomers). It is believed that muscarinic receptors may also be arranged into dimeric or oigomeric complexes, but no systematic experimental evidence exists concerning the direct physical interaction between receptor proteins as its mechanism. We undertook this study to determine whether muscarinic receptors form homomers or a heteromers by direct protein-protein interaction within the same or within different subtypes using a yeast two-hybrid system. Intracellular loops (i1, i2 and i3) and the C-terminal cytoplasmic tails (C) of human muscarinic (Hm) receptor subtypes, Hm1, Hm2 and Hm3, were cloned into the vectors (pB42AD and pLexA) of a two-hybrid system and examined for heteromeric or homodimeric interactions between the cytoplasmic domains. No physical interaction was observed between the intracellular domains of any of the Hm/Hm receptor sets tested. The results of our study suggest that the Hm1, Hm2 and Hm3 receptors do not form dimers or oligomers by interacting directly through either the hydrophilic intracellular domains or the C-terminal tail domains. To further investigate extracellular domain interactions, the N-terminus (N) and extracellular loops (o1 and o2) were also cloned into the two-hybrid vectors. Interactions of Hm2N with Hm2N, Hm2o1, Hm2o2, Hm3N, Hm3o1 or Hm3o2 were examined. The N-terminal domain of Hm2 was found to have no direct interaction with any extracellular domain. From our results, we excluded the possibility of a direct interaction between the muscarinic receptor subtypes (Hm1, Hm2 and Hm3) as a mechanism for homo- or hetero-meric dimerization/oligomerization. On the other hand, it remains a possibility that interaction may occur indirectly or require proper conformation or subunit formation or hydrophobic region involvement.

Comparison of the Food Quality of Hybrid Grouper Epinephelus fuyscoguttatus×E. labnceolatus and Hybrid Longtooth Grouper E. moara×E. lanceolatus (대왕범바리(Epinephelus fuscoguttatus×E. lanceolatus)와 대왕자바리(E. moara×E. lanceolatus)의 식품학적 품질비교)

  • Lee, Woo Jin;An, Byoung Kyu;In, Jung Jin;Han, Hyeong Gu;Park, Jong-Yeon;Bang, In-Chul;Shim, Kil Bo
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.55 no.2
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    • pp.129-136
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    • 2022
  • The nutrient composition of the muscle of the hybrid grouper (Epinephelus fuscoguttatus×E. lanceolatus, HG) and the hybrid longtooth grouper (E. moara×E. lanceolatus, HLG) was measured as a function of body weight in this study. The crude protein and lipid levels in HG were 21.0-21.2 g/100 g and 1.9-3.4 g/100 g, respectively. HLG contained 20.6-20.7 g/100 g protein, and 3.9-5.5 g/100 g crude lipid. The total amino acid contents of HG were 18,829.7-18,980.2 mg/100 g and that of HLG was 17,793.7-19,293.7 mg/100 g. The mean saturated fatty acid content was 0.61-1.09 g/100 g for HG and 1.27-1.77 g/100 g for HLG. Monounsaturated and polyunsaturated fatty acid levels were 0.59-1.02 g/100 g (Monoene), 0.67-1.11 g/100 g (Polyene) in HG, which were lower than the 1.21-1.78 (Monoene), 1.22-1.69 g/100 g (Polyene) found in HLG. The highest mineral content (K and P) of HG was 510.13-517.05 mg/100g and 231.59-247.67 mg/100 g, while that of HLG was 518.81-523.59 mg/100 g and 257.51-248.84 mg/100 g, respectively. In conclusion, there is potential for expanding the commercial utilization of HG and HLG as food resources as they are both nutrient-rich.

The Alpha Subunit of Go Interacts with Brain Specific High Mobility Group Box Containing Protein

  • Park, Jung-Sik;Ghil, Sung-Ho
    • Biomedical Science Letters
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    • v.12 no.4
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    • pp.405-411
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    • 2006
  • Heterotrimeric GTP binding proteins (G proteins) mediate signal transduction generated by neurotransmitter and hormones. Among G-proteins, Go is classified as a member of the Go/Gi family and the most abundant heterotrimeric G protein in brain. Most of the mechanistic analyses on the activation of Go indicated its action to be mediated by the $G{\beta}{\gamma}$ dimer because downstream effectors for its ${\alpha}$ subunit have not been clearly defined. To determine the downstream effectors of alpha subunits of Go ($Go{\alpha}$), we used yeast two-hybrid system to screen $Go{\alpha}$ interacting partners in cDNA library from the human brain. A brain specific high mobility group box containing protein (BHX), A possible transcription factor, was identified as a $Go{\alpha}$ interacting protein. We confirmed interaction between $Go{\alpha}$ and BHX employing in vitro affinity binding assay. Moreover, active form of $Go{\alpha}$ preferentially interacts with BHX than inactive farm. Our findings indicate that $Go{\alpha}$ could modulate gene expression via interaction with BHX during neuronal or brain development.

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Novel calcineurin interacting protein-2: the functional characterization of CNP-2 in Caenorhabditis elegans

  • Xianglan, Cai;Ko, Kyung-Min;Singaravelu, Gunasekaran;Ahnn, Joo-Hong
    • BMB Reports
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    • v.41 no.6
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    • pp.455-460
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    • 2008
  • Calcineurin (Cn) is a serine/threonine phosphatase implicated in a wide variety of biological responses. To identify proteins that mediate Cn signaling pathway effects, we used yeast two-hybrid assays to screen for Cn interacting proteins, discovering a protein encoded by the gene, cnp-2 (Y46G5A.10). Utilizing serially deleted forms of Cn as baits, we demonstrated that the catalytic domain of Cn (TAX-6) binds with CNP-2, and this physical interaction was able to be reconstituted in vitro, supporting our yeast two-hybrid results. cnp-2 is a nematode-specific novel gene found in C. elegans as well as its closest relative, C. briggsae. CNP-2 was strongly expressed in the intestine of C. elegans. To study the function of cnp-2, we performed cnp-2 RNAi knock-down and characterized phenotypes associated with Cn mutants. However, no gross defects were revealed in these RNAi experiments. CNP-2 was proven to be a Cn binding protein; however, its role remains to be elucidated.