• Journal of Life Science
• /
• v.17 no.6 s.86
• /
• pp.783-790
• /
• 2007

G protein coupled receptors (GPCRs) transmit various extracellular signals into the cells. Upon binding of the ligands, conformational changes in the extracellular and/or transmembrane (TM) domains of CPCRs were propagated into the cytoplasmic (CP) domain of the molecule leading to the activation of their cognate heterotrimeric C proteins and kinases. Constitutively active GPCR mutants causing the activation of C Protein signaling even in the absence of ligand binding are of interest for the study of activation mechanism of GPCRs. Two classes of constitutively active mutations, categorized by their effects on the salt bridge between Ell3 and K296, were found in the TM domain of rhodopsin. Opsin mutants containing combinations of the mutations were constructed to study the conformational changes required for the activation of rhodopsin. Rhodopsin chromophore regenerated with 11-cis-retinal showed a thermal stability inversely correlated with its constitutive activity. In contrast, rhodopsin mutants exhibited a binding affinity to an agonist, all-trans-retinal, in a constitutive activity-dependent manner. In order to test whether the conformational changes responsible for the activation of trans-ducin (Gt) are the same as the conformation required for the recognition of rhodopsin kinase, analysis of the mutants were carried out with phosphorylation by rhodopsin kinase. Rhodopsin mutants containing combinations of different classes of the mutations showed a strong synergistic effect on the phosphorylation of the mutants in the dark as similar to that of Gt activation. The results suggest that at least two or three kinds of segmental and independent conformational changes are required for the activation of rhodopsin and the conformational changes responsible for activating rhodopsin kinase and Gt are similar to each other.

• Proceedings of the Korean Society of Embryo Transfer Conference
• /
• 2002.11a
• /
• pp.96-96
• /
• 2002

We examined the role of SR 144528 (N-[-(1S-endo-1,3,,3-trimethyl-bicycle[2, 2, 1 ] heptan-2-y1]-5-(-4-chloro-3-mothyl-phenyl)-(4-methylbenzyl)-pyrazole-3- carboxamide) in the modulation of certain AC isoforms in transiently transfected COS-7 cells. We found that CB2 in COS cells has a constitutive activity, and thus leading to inhibition of AC-V activity even in the absence of agonist. In addition, this constitutive modulation of AC is reversed by SR144528. It is now well established that several G protein-coupled receptors can signal without agonist stimulation(constitutive receptors). Inverse agonists have been shown to inhibit the activity of such constitutive G protein-coupled receptor signaling. Agonist activation of the G$\_$i/o/-coupled peripheral cannabinoid receptor CB2 normally inhibits adenylyl cyclase type V and stimulates adenylyl cyclase type II. Using transfected COS cells, we show here that application of SR144528, an inverse agonist of CB2, leads to a reverse action (stimulation of adenylyl cyclase V and inhibition of adenylyl cyclase II). This inverse agonism of SR144528 is dependent on the temperature, as well as on the concentration of the cDNA of CB2 transfected. Pertussis toxin blocked the regulation of adenylyl cyclase activity by SR 144528.

• Molecules and Cells
• /
• v.27 no.1
• /
• pp.39-45
• /
• 2009

Ligand-dependent or independent oligomerization of receptor protein tyrosine kinase (RPTK) is often an essential step for receptor activation and intracellular signaling. The novel oncogene with kinase-domain (NOK) is a unique RPTK that almost completely lacks an ectodomain, expresses intracellularly and activates constitutively. However, it is unknown whether NOK can form oligomer or what function oligomerization would have. In this study, two NOK deletion mutants were generated by either removing the ectodomain ($NOK{\Delta}ECD$) or including the endodomain (NOK-ICD). Co-immunoprecipitation demonstrated that the transmembrane (TM) domain of NOK was essential for its intermolecular interaction. The results further showed that NOK aggregated more closely as lower order oligomers (the dimer- and trimer-sized) than either deletion mutant did since NOK could be crosslinked by both Sulfo-EGS and formaldehyde, whereas either deletion mutant was only sensitive to Sulfo-EGS. Removing the NOK TM domain (NOK-ICD) not only markedly promoted higher order oligomerization, but also altered the subcellular localization of NOK and dramatically elevated the NOK-mediated constitutive activation of extracellular signal-regulated kinase (ERK). Moreover, NOK-ICD but not NOK or $NOK{\Delta}ECD$ was co-localized with the upstream signaling molecule RAS on cell membrane. Thus, TM-mediated intermolecular contacting may be mainly responsible for the constitutive activation of NOK and contribute to the autoinhibitory effect on RAS/MAPK signaling.

• Advances in materials Research
• /
• v.3 no.4
• /
• pp.217-225
• /
• 2014

In this study, a forging steel alloyed with both Nb and V was used as experimental material and the hot deformation behavior has been studied for this steel by conducting the compressive deformation test at temperature of $900-1150^{\circ}C$ and strain rate of $0.01-0.01s^{-1}$ in a MMS-300 thermo-mechanical simulator. The microstructure evolution, particularly the dynamically recrystallized microstructure, of the experimental steel at elevated temperatures, strain rates and strain levels, was characterized by optical microstructural observation and the constitutive equation in association with the activation energy and Zener-Hollomon parameter. The curves of strain hardening rate versus stress were used to determine the critical strain and peak strain, and their relation was connected with Zener-Hollomon parameter. Under the conditions of processing temperature $900^{\circ}C$ and strain rate $0.01s^{-1}$, the dynamic recrystallization took place and the austenite grain size was refined from $164.5{\mu}m$ to $28.9{\mu}m$.

• Advances in materials Research
• /
• v.1 no.4
• /
• pp.245-268
• /
• 2012

Results of isothermal torsional oscillation tests are reported on melts of linear low density polyethylene and isotactic polypropylene. Prior to rheological tests, specimens were annealed at various temperatures ranging from $T_a$ = 180 to $310^{\circ}C$ for various amounts of time (from 30 to 120 min). Thermal treatment induced degradation of the melts and caused pronounced decreases in their molecular weights. With reference to the concept of transient networks, constitutive equations are developed for the viscoelastic response of polymer melts. A melt is treated as an equivalent network of strands bridged by junctions (entanglements and physical cross-links). The time-dependent response of the network is modelled as separation of active strands from and merging of dangling strands with temporary nodes. The stress-strain relations involve three adjustable parameters (the instantaneous shear modulus, the average activation energy for detachment of active strands, and the standard deviation of activation energies) that are determined by matching the dependencies of storage and loss moduli on frequency of oscillations. Good agreement is demonstrated between the experimental data and the results of numerical simulation. The study focuses on the effect of molecular weight of polymer melts on the material constants in the constitutive equations.

• Journal of Life Science
• /
• v.21 no.4
• /
• pp.481-485
• /
• 2011

T0901317 is a potent synthetic ligand for liver X receptor (LXR, NR1H2/3), a member of the nuclear receptor superfamily that functions as a transcription factor. However, T0901317 has been also reported to modulate the activity at least four other nuclear receptors (NRs), acting as agonists for farnesoid X receptor (FXR, NR1H4) and pregnane X receptor (PXR, NR1I2) and as antagonists for androgen receptor (AR, NR3C4) and retinoid-related orphan receptor-${\alpha}$ (ROR-${\alpha}$, NR1F1). We report here that T0901317 can also function as an inhibitor for constitutive androstane receptor (CAR, NR1I3). Since CAR is a major player of xenobiotic and cholesterol metabolism in the liver, along with PXR, FXR and LXR, which are reported to be regulated by T0901317, this further complicates the interpretation of potential results with T0901317 in liver cells.

• Proceedings of the Korean Society for Technology of Plasticity Conference
• /
• 2003.05a
• /
• pp.57-60
• /
• 2003

The purpose of this study is to investigate the high temperature deformation behavior of Ti-6Al-4V alloy and to predict the final microstructure under given forming conditions. Equiaxed and widmanstatten of Ti-6Al-4V alloys were prepared as initial microstructure and the compression tests were performed to obtain the flow curves at high temperatures (700∼1100$^{\circ}C$) and various strain rates (10$\^$-4/∼10$^2$/s). Form the results of compression test various parameters such as strain rate sensitivity (m) and activation energy (Q) were calculated and used to establish constitutive equations. To predict the final microstructure after forming, finite element analysis was performed considering the microstructural parameters such as the grain size and the volume fraction of second phase.

• Animal cells and systems
• /
• v.5 no.3
• /
• pp.247-251
• /
• 2001

Differentiating HiB5 cells, a rat hippocampal cell line, expressed neuregulins and showed constitutive activation of a neuregulin receptor, ErbB2, suggesting development of a neuregulin autocrine loop. RT-PCR analyses indicated that HiB5 cells produced SMDF and NDF, but not GGF, during the differentiation. None of neuregulin isoforms were detected in proliferating HiB5 cells. The neuregulins in HiBS cells, at least in part, are the $\beta$-isoforms of which the most of neuronal neuregulin isoforms are. The expression of SMDF and NDF was enhanced by PDGF and bFGF that promote cell survival and differentiation, suggesting a close relationship between the synthesis of neuregulins and the differentiation process. HiB5 cells have ErbB2 and ErbB4, but not ErbB3 receptors. Constitutive tyrosine phosphorylation of ErbB2 was detected in HiB5 cells that had not been exposed to exogenous GGF.

• Transactions of Materials Processing
• /
• v.12 no.4
• /
• pp.290-295
• /
• 2003

High temperature deformation behavior and prediction of final microstructure after forming of Ti-6Al-4V alloy were investigated in this study. Equiaxed and Widmanstatten microstructures of Ti-6Al-4V alloys were prepared as initial microstructures and compression tests were performed to obtain the flow curves at high temperatures (700∼110$0^{\circ}C$) and various strain rates (10$^{-4}$ ∼10$^2$/s). From the results of compression test, strain rate sensitivity (m) and activation energy (Q) were calculated and used to establish constitutive equation. To predict the final microstructure after farming, finite element analysis was performed considering the microstructural parameters such as grain size and volume fraction of second phase.

• Oncology Letters
• /
• v.41 no.5
• /
• pp.3119-3126
• /
• 2019

Redd1 is a stress response protein that functions as a repressor of mTORC1, a central regulator of protein translation, resulting in the inhibition of cell growth and metabolism. However, paradoxically, high Redd1 expression favors cancer progression and generates resistance to cancer therapy. Herein, we revealed that constitutive overexpression of Redd1 induced HSP27 and HSP70 expression in lung cancer cells. The expression of Redd1, HSP27 and HSP70 was highly increased in lung cancer tissues compared with that in normal lung tissues. Inhibition of HSP27 or HSP70 suppressed AKT phosphorylation, which was induced by constitutive overexpression of Redd1 and enhanced the inhibitory effects on viability of Redd1-overexpressing cells. Inhibition of AKT phosphorylation resulted in a decrease of HSP27 and HSP70 expression in Redd1-overexpressing cells. These data indicated that HSPs and AKT in Redd1-overexpressing cells positively regulated the function and expression of each other and were involved in lung cancer cell survival. Knockdown of HSP27, HSP70 or AKT enhanced ionizing radiation (IR) sensitivity, particularly in lung cancer cells in which Redd1 was stably overexpressed. Collectively, constitutive overexpression of Redd1 led to HSP27 and HSP70 induction and AKT activation, which were involved in lung cancer cell survival and resistance to IR, suggesting that Redd1 may be used as a therapeutic target for lung cancer.