• Title/Summary/Keyword: Binding Mechanism

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A Ser/Thr Specific Protein Kinase Activates the Mouse Rantes Gene after Lipolpolysaccharide STimulation

  • Kim, Youn-Uck;Kim, Youn-Hwoan;An, Duek -Jun;Kwon, Hyuk-Chu
    • Journal of Microbiology
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    • v.39 no.4
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    • pp.314-320
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    • 2001
  • Macrophages stimulated by lipopolysaccharide(LPS) from gram negative bacteria undergo activation of a group of immediate early genes including Rantes. The mouse Rantes gene promoter region contains an LPS rsponsive element(LPE) We detected 3 specific bands termed B1, B2 and 3 formed by the interaction of the LPE and proteins found in LPS-stimulated RAW 367.7 cells. An additional band B4 was determined to be an Ap-1 binding protein. The B1 band appears within 1 hour of LPS nuclear extracts from LPS-stimulation, and this protein kinase enhances B1 and formation. The B1 band can be converted to band B2/B3 by adding specific heparin column fraction purified Ser/Thr specific protein phosphatases PP-1 and PP-2A can stimulate the same conversion to about the same extent. Thus, the formation of the LRE sequence binding complex appears to be regulated by Ser/Thr protein kinase and one or more Ser/Thr specific phosphatases. At least four proteins are involved in the trgulation of the LRE-dependent Rants experssion: two binding factors that bind directly to the target sequences. and two factors that control their binding. The future purification and characterization of these binding pro-teins will reveal in detail the mechanism of Rantes gene activation after LPS stimulation.

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Change of hCG Binding Capacity on the Granulosa Cell of Porcine Ovary During Follicular Atresia (돼지 난소내 여포의 폐쇄과정중 과립세포의 결합능의 변화)

  • Chang, Choul-Soo;Lee, Chang-Joo;Yoon, Yong-DaI;Kim, Moon-Kyoo
    • Clinical and Experimental Reproductive Medicine
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    • v.13 no.1
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    • pp.29-38
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    • 1986
  • In order to study the mechanism of follicular atresia, follicles were classified into the normal groups and the atretic ones, according to the criteria with or without corpus luteum, size of follicles, vascularization, status of granulose cells and the hypertrophy of theca layers in the porcine ovary. To estimate the binding capacity of human chorionic gonadotropin (hCG) receptor on the granulosa cells during atresia, hCG were iodinated by the chloramine-T method and then purified through the column chromatography. The concentration" of hCG receptor in each group were measured by the hCG receptor binding assay. Binding capacity in large normal follicles were 1.16%, but atretic ones were 0.45%. But in medium and small follicles (below 6mm in diameter), the binding capacity in normal follicles were 0.09%, but atretic ones were 0.05%, which was lower than those of large follicles. The present ( ) that the concentrations of hCG receptors on granulosa cells is decreased when the follicles become atretic and be used as a sort of creteria for the identification of follicles atresia.

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Kinetics and Mechanism of Mutant O-acetylserine Sulfhydrylase-A (C43S) from Salmonella typhimurium LT-2

  • Yoon, Moon-Young
    • BMB Reports
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    • v.29 no.3
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    • pp.210-214
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    • 1996
  • The pH dependence of the kinetic parameters of mutant O-acetylserine sulfhydrylase (OASS) from Salmonella typhimurium LT-2 has been determined in order to obtain information on the chemical mechanism. The initial velocity pattern obtained by varying the concentrations of OAS at several fixed concentrations of TNB, shows an intersection on the left of the ordinate at pH 7.0, indicating that the kinetic mechanism is a sequential mechanism in which substrate inhibition by OAS is observed while the wild type enzyme showed a ping pong mechanism. The values of $V/E_t$, $V/K_{OAS}E_{t}$ and $V/K_{TNB}E_{t}$ decreased by about 68%, 14% and 16% as compared with the wild type enzyme. The $V/K_{OAS}E_{t}$ is a pK of 6.5 on the acid side of the pH profile, and the $V/K_{TNB}$ is pH independent. As compared with the wild type enzyme, the pKs in the V/K profiles are shifted, reflecting that binding of the cofactor in free E:OAS is less asymmetric.

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A Study on the Nylon Wire Holding and Release Mechanism for Cube Satellites by Applying Constant Holding

  • Koo, KeonWoo
    • Journal of Aerospace System Engineering
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    • v.15 no.4
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    • pp.1-6
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    • 2021
  • The non-explosive holding and release mechanism is used to prevent damage to the mission component caused by explosives when the deployment structure for Cube Satellites is separated. However, among the several types available, the non-explosive holding and release mechanism system using nylon wires depend on the nylon wire knot method and tightening power of the worker. Therefore, in this study, we conducted experiments with the operation of a new holding and release mechanism system by conceptualizing the Boa System Dial, which can provide a constant tightening force regardless of worker proficiency and deploying a imitational solar panel. In this study, the process of binding and unbinding with constant tension was recorded while applying the novel non-explosive holding and release mechanism using the Boa System Dial proposed. In addition, required advances are indicated for the application of the proposed system to actual Cube Satellites.

Curcumin Derivatives Inhibit the Formation of Jun-Fos-DNA Complex Independently of their Conserved Cysteine Residues

  • Park, Chi-Hoon;Lee, Ju-Hyung;Yang, Chul-Hak
    • BMB Reports
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    • v.38 no.4
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    • pp.474-480
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    • 2005
  • Curcumin, a major active component of turmeric, has been identified as an inhibitor of the transcriptional activity of activator protein-1 (AP-1). Recently, it was also found that curcumin and synthetic curcumin derivatives can inhibit the binding of Jun-Fos, which are the members of the AP-1 family, to DNA. However, the mechanism of this inhibition by curcumin and its derivatives was not disclosed. Since the binding of Jun-Fos dimer to DNA can be modulated by redox control involving conserved cysteine residues, we studied whether curcumin and its derivatives inhibit Jun-Fos DNA binding activity via these residues. However, the inhibitory mechanism of curcumin and its derivatives, unlike that of other Jun-Fos inhibitors, was found to be independent of these conserved cysteine residues. In addition, we investigated whether curcumin derivatives can inhibit AP-1 transcriptional activity in vivo using a luciferase assay. We found that, among the curcumin derivatives examined, only inhibitors shown to inhibit the binding of Jun-Fos to DNA by Electrophoretic Mobility Shift Assay (EMSA) inhibited AP-1 transcriptional activity in vivo. Moreover, RT-PCR revealed that curcumin derivatives, like curcumin, downregulated c-jun mRNA in JB6 cells. These results suggest that the suppression of the formation of DNA-Jun-Fos complex is the main cause of reduced AP-1 transcriptional activity by curcuminoids, and that EMSA is a suitable tool for identifying inhibitors of transcriptional activation.

Sleep Promoting Effect of Luteolin in Mice via Adenosine A1 and A2A Receptors

  • Kim, Tae-Ho;Custodio, Raly James;Cheong, Jae Hoon;Kim, Hee Jin;Jung, Yi-Sook
    • Biomolecules & Therapeutics
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    • v.27 no.6
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    • pp.584-590
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    • 2019
  • Luteolin, a widespread flavonoid, has been known to have neuroprotective activity against various neurologic diseases such as epilepsy, and Alzheimer's disease. However, little information is available regarding the hypnotic effect of luteolin. In this study, we evaluated the hypnotic effect of luteolin and its underlying mechanism. In pentobarbital-induced sleeping mice model, luteolin (1, and 3 mg/kg, p.o.) decreased sleep latency and increased the total sleep time. Through electroencephalogram (EEG) and electromyogram (EMG) recording, we demonstrated that luteolin increased non-rapid eye movement (NREM) sleep time and decreased wake time. To evaluate the underlying mechanism, we examined the effects of various pharmacological antagonists on the hypnotic effect of luteolin. The hypnotic effect of 3 mg/kg of luteolin was not affected by flumazenil, a GABAA receptorbenzodiazepine (GABAAR-BDZ) binding site antagonist, and bicuculine, a GABAAR-GABA binding site antagonist. On the other hand, the hypnotic effect of 3 mg/kg of luteolin was almost completely blocked by caffeine, an antagonist for both adenosine A1 and A2A receptor (A1R and A2AR), 8-Cyclopentyl-1,3-dipropylxanthine (DPCPX), an A1R antagonist, and SCH-58261, an A2AR antagonist. From the binding affinity assay, we have found that luteolin significantly binds to not only A1R but also A2AR with $IC_{50}$ of 1.19, $0.84{\mu}g/kg$, respectively. However, luteolin did not bind to either BDZ-receptor or GABAAR. From these results, it has been suggested that luteolin has hypnotic efficacy through A1R and A2AR binding.

Analysis on Adsorption Rate & Mechanism on Chloride Adsorption Behavior with Cement Hydrates (시멘트 수화물의 염소이온 흡착거동에 따른 메커니즘 및 해석기법)

  • Yoon, In-Seok
    • Journal of the Korea Concrete Institute
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    • v.27 no.1
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    • pp.85-92
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    • 2015
  • The chloride ions, responsible for the initiation of the corrosion mechanism, intrude from the external medium into the concrete. A part of the intruding chloride ions will be retained by the hydration products of the binder in concrete, either through chemical adsorption or by physical adsorption. Since the hydration products of cement are responsible for the chloride binding in concrete, this study focused on the chloride binding in individual hydrate. The purpose of this study is to explore the time dependant behaviors of chloride ions adsorption with cement hydrates, focused on its mechanism. AFt phase and CH phase were not able to absorb chloride ion, however, C-S-H phase and AFm phase had a significant chloride adsorption capacity. In particular, AFm phase showed a chemical adsorption with slow rate in 40 days, while C-S-H phase showed binding behaviors with 3 stages including momentary physical adsorption, physico-chemical adsorption, and chemical adsorption. Based on the results, this study suggested theoretical approach to depict chloride adsorption behavior with elapsed time of C-S-H phase and AFm phase effectively. It is believed that the approach suggested in this study can provide us with a good solution to understand the mechanism on chloride adsorption with hydrates and to calculate a rate of chloride penetration with original source of chloride ions, for example, marine sand at initial time or sea water penetration later on.

Recent Advances in Structural Studies of Antifreeze Proteins (구조 생물학을 이용한 Antifreeze protein의 최근 연구동향)

  • Lee, Jun-Hyuck;Lee, Sung-Gu;Kim, Hak-Jun
    • Ocean and Polar Research
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    • v.33 no.2
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    • pp.159-169
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    • 2011
  • Antifreeze proteins (AFPs) have ice binding affinity, depress freezing temperature and inhibit ice recystallization which protect cellular membranes in polar organisms. Recent structural studies of antifreeze proteins have significantly expanded our understanding of the structure-function relationship and ice crystal growth inhibition. Although AFPs (Type I-IV AFP from fish, insect AFP and Plant AFP) have completely different fold and no sequence homology, they share a common feature of their surface area for ice binding property. The conserved ice-binding sites are relatively flat and hydrophobic. For example, Type I AFP has an amphipathic, single ${\alpha}$-helix and has regularly spaced Thr-Ala residues which make direct interaction with oxygen atoms of ice crystals. Unlike Type I AFP, Type II and III AFP are compact globular proteins that contain a flat ice-binding patch on the surface. Type II and Type III AFP show a remarkable structural similarity with the sugar binding lectin protein and C-terminal domain of sialic acid synthase, respectively. Type IV is assumed to form a four-helix bundle which has sequence similarity with apolipoprotein. The results of our modeling suggest an ice-binding induced structural change of Type IV AFP. Insect AFP has ${\beta}$-helical structure with a regular array of Thr-X-Thr motif. Threonine residues of each Thr-X-Thr motif fit well into the ice crystal lattice and provide a good surface-surface complementarity. This review focuses on the structural characteristics and details of the ice-binding mechanism of antifreeze proteins.

HCBP6 upregulates human SREBP1c expression by binding to C/EBPβ-binding site in the SREBP1c promoter

  • Yang, Xueliang;Han, Ming;Liu, Shunai;Yuan, Xiaoxue;Liu, Xiaojing;Feng, Shenghu;Zhou, Li;Li, Yaru;Lu, Hongping;Cheng, Jun;Lin, Shumei
    • BMB Reports
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    • v.51 no.1
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    • pp.33-38
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    • 2018
  • Sterol regulatory element-binding protein-1c (SREBP1c) plays an important role in triglyceride (TG) homeostasis. Although our previous study showed that hepatitis C virus core-binding protein 6 (HCBP6) regulates SREBP1c expression to maintain intracellular TG homeostasis, the mechanism underlying this regulation is unclear. In the present study, we found that HCBP6 increased intracellular TG levels by upregulating SREBP1c expression. HCBP6 increased SREBP1c transcription by directly binding to the SREBP1c promoter (at the -139- to +359-bp region). Moreover, we observed that HCBP6 interacted with $C/EBP{\beta}-binding$ site in the SREBP1c promoter both in vitro and in vivo. These results indicate that HCBP6 upregulates human SREBP1c expression by binding to the $C/EBP{\beta}-binding$ site in the SREBP1c promoter.

Interaction of Antihistaminics with Muscarinic Receptor(I) -Action on the cardiac muscarinic receptor- (항(抗) Histamine제(劑)와 Muscarinic Receptor와의 상호작용(相互作用)(I) -심장(心臟) muscarinic receptor에 대한 작용(作用)-)

  • Lee, Shin-Woong;Park, Yeung-Joo;Lee, Jeung-Soo;Ha, Kwang-Won;Jin, Kap-Duck
    • YAKHAK HOEJI
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    • v.32 no.2
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    • pp.101-111
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    • 1988
  • $[^3H]$ Quinuclidinyl benzilate(QNB) binding assays were performed in the dog ventricular sarcolemma fraction enriched approx. 32-fold in sarcolemma compared to the starting homogenate to elucidate the effect of antihistaminics on cardiac muscarinic receptor. Chlorpheniramine(CHP) inhibited specific binding of $[^3H]$QNB and delayed the equilibrium binding. The rate constants at $37^{\circ}C$ for formation and dissociation of the QNB receptor complex were $0.38{\times}10^9\;M^{-1}$ and $1.6{\times}10^{-2}\;min^{-1}$, respectively. The mean value for the dissociation constant from the pairs of the rate constants was 43. 2 pM and this value was similar to the value(44.8pM) determined from Scatchard analysis. CHP decreased association rate constant, indicating increase in $K_D$ value. Decrease in affinity without affecting the binding site concentration$(B_{max})$ for $[^3H]$QNB binding by CHP was also demonstrated by Scatchard analysis. $K_i$ values for $H_i$-blockers that inhibited specific $[^3H]$QNB binding were $0.02{\sim}4.8{\mu}M$. Cimetidine with $K_i$ value of $230{\mu}M$, however, was ineffective in displacing $[^3H]$QNB binding at concentration of $50{\mu}M$. The Hill coefficient for $H_1$-blockers were about one. The results indicate that $H_1$-antihistaminics inhibit $[^3H]$ QNB binding by interaction with myocardiac muscarinic cholinergic receptor and anticholinergic side effects of these drugs are mainly due to this receptor blocking mechanism.

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