• 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.

• Journal of Life Science
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• v.22 no.12
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• pp.1608-1613
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• 2012

A conventional kinesin, KIF5/Kinesin-I, transports various cargoes along the microtubule through interaction between its light chain subunit and the cargoes. Kinesin light chains (KLCs) interact with many different cargoes using their tetratricopeptide repeat (TPR) domain, but the mechanism underlying recognition and binding of a specific cargo has not yet been completely elucidated. We used the yeast two-hybrid assay to identify proteins that interact with the TPR domain of KLC1. We found an interaction between the TPR domain of KLC1 and an amyloid precursor protein (APP)-binding protein PAT1 (protein interacting with APP tail 1). The yeast two-hybrid assay demonstrated that the TPR domain-containing region of KLC1 mediated binding to the C-terminal tail region of PAT1. PAT1 also bound to KLC2 but not to kinesin heavy chains (KIF5A, KIF5B, and KIF5C) in the yeast two-hybrid assay. These protein-protein interactions were also observed in the glutathione S-transferase (GST) pull-down assay and by co-immunoprecipitation. Anti-PAT1 antibody as well as anti-APP anti-body co-immunoprecipitated KLC and KHCs associated with PAT1 from mouse brain extracts. These results suggest that PAT1 could mediate interactions between Kinesin-I and APP containing vesicles.

• Journal of Life Science
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• v.20 no.7
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• pp.1005-1011
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• 2010

$\gamma$-aminobutyric acid (GABA) is the major inhibitory neurotransmitter in the central nervous system. GABA transporters (GATs) control extracellular GABA levels by reuptake of released GABA from the synaptic cleft. However, how GATs are regulated has not yet been elucidated. Here, we used the yeast two-hybrid system to identify the specific binding protein(s) that interacts with the carboxyl (C)-terminal region of GAT1, the major isoform in the brain and find a specific interaction with the ubiquitin-specific protease 14 (Usp14), a deubiquitinating enzyme. Usp14 protein bound to the tail region of GAT1 and GAT2 but not to other GAT members in the yeast two-hybrid assay. The C-terminal region of Usp14 is essential for interaction with GAT1. In addition, these proteins showed specific interactions in the glutathione S-transferase (GST) pull-down assay. An antibody to GAT1 specifically co-immunoprecipitated Usp14 from mouse brain extracts. These results suggest that Usp14 may regulate the number of GAT1 at the cell surface.

• Journal of Life Science
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• v.19 no.7
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• pp.968-975
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• 2009

The purpose of the present study was to examine effects of different exercise training modes (Aerobic Training, Resistance Training) on exercise specificity and transability. The tested subjects, composed of 10 healthy males without known family history or medical illnesses, were divided into two groups: Aerobic Training Group (ATG; n=5) and Resistance Training Group (RTG; n=5). An aerobic training program, based on maximum oxygen consumption rates taken during standard testing, was conducted in 60 minute sessions 3 times a week, and the Heart Rate Reserve (HRR) at 70% of maximum oxygen consumption rate was measured the using Polar. In the weight training program, based on repetition maximum rate (1-RM) taken during standard testing, the weight at 70% of such rates was measured during 60 minute sessions of 7 categories of exercise (Bench press, Leg press, Squat, Shoulder press, Arm curt Lat pull down, Triceps pull down), conducted 3 times a week. The data collected from this research were calculated to obtain average and differences compared to standards using an SPSS 11.0 statistics package. In conclusion, increase in V0$_{2max}$ and production of NO$_x$ (NO$_2$/NO$_3$), reduction of %fat, MAPwere shown effective in aerobic training and in different exercise tests, and aerobic testing within the aerobic training group (ATG) was shown to be more effective. In contrast, resistance training was shown to be more effective for the reduction of CK and LDH, and even in different tests, the resistance test within the resistance training group (RTG) showed to be more effective. Exercise specificity also significantly increased in both groups (ATG, RTG). but there was no significant difference in transability in both groups (ATG, RTG).

• Molecules and Cells
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• v.37 no.3
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• pp.264-269
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• 2014

The molecular interaction between tumor suppressor p53 and the anti-apoptotic Bcl-2 family proteins plays an essential role in the transcription-independent apoptotic pathway of p53. In this study, we investigated the binding of p53 DNA-binding domain (p53DBD) with the anti-apoptotic Bcl-2 family proteins, Bcl-w, Mcl-1, and Bcl-2, using GST pull-down assay and NMR spectroscopy. The GST pull-down assays and NMR experiments demonstrated the direct binding of the p53DBD with Bcl-w, Mcl-1, and Bcl-2. Further, NMR chemical shift perturbation data showed that Bcl-w and Mcl-1 bind to the positively charged DNA-binding surface of p53DBD. Noticeably, the refined structural models of the complexes between p53DBD and Bcl-w, Mcl-1, and Bcl-2 showed that the binding mode of p53DBD is highly conserved among the anti-apoptotic Bcl-2 family proteins. Furthermore, the chemical shift perturbations on Bcl-w, Mcl-1, and Bcl-2 induced by p53DBD binding occurred not only at the p53DBD-binding acidic region but also at the BH3 peptide-binding pocket, which suggests an allosteric conformational change similar to that observed in Bcl-$X_L$. Taken altogether, our results revealed a structural basis for a conserved binding mechanism between p53DBD and the anti-apoptotic Bcl-2 family proteins, which shed light on to the molecular understanding of the transcription-independent apoptosis pathway of p53.

• Journal of the Korea Academia-Industrial cooperation Society
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• v.17 no.10
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• pp.319-330
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• 2016

Recently, 3D printing technology has been applied to make a concept model and working mockup of an industrial application. On the other hand, this technology has limited applications in industrial products due to the materials and reliability of the 3D printed product. In this study, the components of a full cord switch module are proposed as a case of a 3D printed component that can be used as a substitute for a short period. These are hub-driven and lever lockup components that have the structural characteristics of breaking down frequently in the emergency operating status. To ensure the structural strength for a substitute period, research of structure optimization was performed because 3D printing technology has a limitation in the materials used. After optimizing the structure variables of the hub-driven component, reasonable results can be drawn in that the safety factors of the left and right switching mode are 1.243 (${\Delta}153.67%$) and 3.156 (${\Delta}404.96%$). The lever lockup component has a structural weak point that can break down easily on the lockup-part because of a cantilever shape and bending moment. The rib structure was applied to decrease the deflection. In addition, optimization of the structural variables was performed, showing a safety factor of 7.52(${\Delta}26%$).

• Asian Pacific Journal of Cancer Prevention
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• v.14 no.11
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• pp.6305-6309
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• 2013

Background: TIAM2, a Rac guanine nucleotide exchange factor, is closely associated with cell adherence and migration. Here, we aimed to investigate the role of TIAM2 in non-small cell lung cancer (NSCLC) cells. Materials and Methods: A small interference RNA (siRNA) was introduced to silence the expression of TIAM2. Invasion and motility assays were then performed to assess the invasion and motility potential of NSCLC cells. GST-pull down assays were used to detect activation of Rac1. Results: TIAM2 was highly expressed in NSCLC cells. Knockdown of TIAM2 inhibited the invasion and motility, and suppressed activation of Rac1. Further experiments demonstrated that knockdown of TIAM2 could up-regulate the expression of E-cadherin, and down-regulate the expression of MMP-3, Twist and Snail. Conclusions: Our data suggest that TIAM2 can promote invasion and motility of NSCLC cells. Activation of Rac1 and regulation of some EMT/invasion-related genes may be involved in the underlying processes.

• Journal of Korean Society of Industrial and Systems Engineering
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• v.28 no.3
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• pp.75-86
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• 2005

To investigate the fatigue characteristic of upper limbs, this study analyzed RMS(root mean square) and MPF(mean power frequency) value between initial and terminal stages of each experiment condition. And the effect of intermittent endurance time was evaluated using the Borg's CR10 value that was measured for the parts of upper limb. According to the results of ANOVA on RMS value, there were significant difference on the %MVC about push, pull, and down force exertion. Particularly the ANOVA of up force exertion was significant difference on shoulder flexion, elbow flexion and rest time as well as %MVC. The results of ANOVA for MPF value were significant difference on the %MVC in regard of the push and up force exertion. In case of up force exertion, MPF value tended to shift low frequency at all of the experiment conditions. According to the analysis of duty cycle, RMS value considerably increased over 50% duty cycle and as the %MVC increased, the duty cycle affected the increase of RMS value. MPF value for up and down force exertion decreased at 33%, 50% and 67% duty cycle for all of %MVC. Borg CR10 value of hand and forearm were below the 3-point to the 40% of endurance time at 30%MVC and to the 20% of endurance time at 50%MVC with the exception of up force exertion. But Borg CR10 values of upper arm and shoulder at up force exertion were more than 3-point to the 20% of endurance time at 30%MVC and in the start point of endurance time at 50%MVC.

• Animal cells and systems
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• v.6 no.3
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• pp.277-282
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• 2002

Topoisomearse II is an essential enzyme in all organisms with several independent roles in DNA metabolism. Recently, it has been demonstrated that the C-terminal region of topoisomerases II is associated with hetero-logous protein-protein interactions in human and yeast. In this study, we identified that RTP1, a rat homologue of EIA binding protein BS69, is another topoisomerae II interacting protein by yeast two-hybrid screening. RTP1 has an E1A-binding domain and a MYND motif, which are known to be required for transcriptional regulation by binding to other proteins and interaction with the leucine zipper motif of topoisomerase II. The physical interaction between RTP1 and topoisomerase ll$\alpha$ was examined by GST pull-down assay in vitro. The expression level of RTP1 peaks in S phase as that of topoisomerase ll$\alpha$. These results suggest that the interaction between topoisomerase ll$\alpha$ and RTP1 might play an important role in regulating the transcription of genes involved in DNA metabolism in higher eukaryotes.

• Molecules and Cells
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• v.24 no.3
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• pp.372-377
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• 2007

The orphan nuclear receptor, SF-1, plays a pivotal role in the development and differentiation of the endocrine and reproductive systems, and also regulates the transcription of a host of genes, including those encoding several steroidogenic enzymes and gonadotropins. We found that a previously unidentified repressor, EID-1, is an SF-1-interacting protein that inhibits the transactivation of SF-1. A transient transfection assay revealed that EID-1 inhibits SF-1, but not LRH-1, $ERR{\gamma}$, or mCAR. Using the yeast two hybrid and GST pull-down assays, we determined that EID-1 interacted strongly with SF-1. In addition, it colocalized with SF-1 in mammalian cells and interacted specifically with the AF-2 domain of SF-1, competing with SRC-1 to inhibit SF-1 transactivation. EID-1 is expressed in the mouse testis, and its expression decreases during testis development. The results of the present study suggest that EID-1 can act as a repressor, regulating the function of SF-1.