• Title/Summary/Keyword: Binding state

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A Comparative Study of Protein Profiles in Porcine Fetus Fibroblast Cells with Different Confluence States

  • Han, Rong-Xun;Kim, Hong-Rye;Diao, Yunfei;Kim, Myung-Youn;Park, Chang-Sik;Jin, Dong-Il
    • Reproductive and Developmental Biology
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    • v.33 no.4
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    • pp.243-248
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    • 2009
  • To examine the differential expression of proteins during the cycling (70~80% confluences) and G0/G1 (full confluences) phases in porcine fetal fibroblast cells, we used a global proteomics approach by 2-D gel electrophoresis (2-DE) and MALDI-TOF-MS. Cycling cell were harvested at approximately 70% to 80% confluent state while cells in G0/G1 phase were recovered after maintenance of a confluent state for 48 hr. Cellular proteins with isoelectric points ranging between 3.0~10.0, were analyzed by 2-DE with 2 replicates of each sample. A total of approximately 700 spots were detected by 2.D gels stained with Coomassie brilliant blue. On comparing the cell samples obtained from the cycling and G0/G1 phases, a total of 13 spots were identified as differentially expressed proteins, of which 8 spots were up-regulated in the cycling cell and 5 were up-regulated in the G0/G1 phase. Differentially expressed proteins included K3 keratin, similar to serine protease 23 precursor, protein disulfide-isomerase A3, microsomal protease ER-60, alpha-actinin-2, and heat-shock protein 90 beta. The identified proteins were grouped on the basis of their basic functions such as molecular binding, catabolic, cell growth, and transcription regulatory proteins. Our results show expression profiles of key proteins in porcine fetal fibroblast cells during different cell cycle status.

Object-Oriented Software Interaction Test Techniques using Design/CPN (Design/CPN을 이용한 객체지향 소프트웨어 상호작용 테스트 기법)

  • Li, Ren-Ge;Koo, Yeon-Seol
    • The KIPS Transactions:PartD
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    • v.11D no.3
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    • pp.649-658
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    • 2004
  • An object-oriented system is organized by a set of interacting objects and the system behavior is represented by the cooperating interaction bet ween objects. The characteristics of object-oriented software. such as inheritance and polymorphism, increase the difficulty of the object-oriented software testing. At running time of a program, one call from a member function can bind to other member functions because of the dynamic characteristics such as concurrence, dynamic binding and interaction. Therefore, there need the research about considering the characteristics of object-oriented software and concurrently testing the interaction between objects. In this paper, we propose the techniques as follows. First, we construct a flattened state chart diagram by considering the inheritance and polymorphism. Next, we model the system with CPN(Colored Petri Net) that usually is applying the system modeling and simulation. Last, we propose a test case generation techniques for testing the interaction between objects in object-oriented software by applying a Design/CPN tool.

Effects of 3,3',4,4',5-pentachlorobiphenyl on human Kv1.3 and Kv1.5 channels

  • Kim, Jong-Hui;Hwang, Soobeen;Park, Seo-in;Jo, Su-Hyun
    • International Journal of Oral Biology
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    • v.44 no.3
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    • pp.115-123
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    • 2019
  • Among the environmental chemicals that may be able to disrupt the endocrine systems of animals and humans are polychlorinated biphenyls (PCBs), a chemical class of considerable concern. PCB consists of two six-carbon rings linked by a single carbon bond, and theoretically, 209 congeners can form, depending on the number of chlorines and their location on the biphenyl rings. Furthermore, 3,3',4,4',5-pentachlorobiphenyl (PCB126) exposure also increases nitric oxide production and nuclear factor kappa-light-chain-enhancer of activated B cells binding activity in chondrocytes, thus contributing as an initiator of chondrocyte apoptosis and resulting in thymic atrophy and immunosuppression. This study identified whether cardiac and immune abnormalities from PCB126 were caused by the Kv1.3 and Kv1.5 channels. PCB126 did not affect either the steady-state current or peak current of the Kv1.3 and Kv1.5 channels. However, PCB126 right-shifted the steady-state activation curves of human Kv1.3 channels. These results suggest that PCBs can affect the heart in a way that does not block voltage-dependent potassium channels including Kv1.3 and Kv1.5 directly.

Design and Fabrication of Ka-band High Power and Low Loss Waveguide Combiner (Ka 대역 고출력 저손실 도파관 결합기 설계 및 제작)

  • Kim, Hyo-Chul;Cho, Heung-Rae;Lee, Ju-Heun;Lee, Deok-Jae;An, Se-Hwan;Lee, Man-Hee;Joo, Ji-Han;Kwon, Jun-Beom;Jeong, Hae-Chang;Kim, So-Su
    • The Journal of the Institute of Internet, Broadcasting and Communication
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    • v.21 no.3
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    • pp.35-42
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    • 2021
  • The research of amplifier have been actively conducted to replace the Traveling Wave Tube Amplifier (TWTA) in the mmWave. For Solid State Power Amplifiers (SSPA), which combine semiconductor-type devices to obtain high output, Low-loss, high-efficiency combination techniques are required to meet the required output as the output of a single relatively low device is relatively low. In this paper, we design and produce an 8-way waveguide combiner and a reflective loss of more than 20dB and a binding efficiency of 85% or more were identified. Field analysis calculates the critical power inside the combiner. It secured stable Power Ratings and built-in coupler for power monitoring to achieve miniaturization and light weight.

Functional Characterization of Drosophila melanogaster CYP6A8 Fatty Acid Hydroxylase

  • Sang-A Lee;Vitchan Kim;Byoungyun Choi;Hyein Lee;Young-Jin Chun;Kyoung Sang Cho;Donghak Kim
    • Biomolecules & Therapeutics
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    • v.31 no.1
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    • pp.82-88
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    • 2023
  • Genomic analysis indicated that the genome of Drosophila melanogaster contains more than 80 cytochrome P450 genes. To date, the enzymatic activity of these P450s has not been extensively studied. Here, the biochemical properties of CYP6A8 were characterized. CYP6A8 was cloned into the pCW vector, and its recombinant enzyme was expressed in Escherichia coli and purified using Ni2+-nitrilotriacetate affinity chromatography. Its expression level was approximately 130 nmol per liter of culture. Purified CYP6A8 exhibited a low-spin state in the absolute spectra of the ferric forms. Binding titration analysis indicated that lauric acid and capric acid produced type I spectral changes, with Kd values 28 ± 4 and 144 ± 20 µM, respectively. Ultra-performance liquid chromatography-mass spectrometry analysis showed that the oxidation reaction of lauric acid produced (ω-1)-hydroxylated lauric acid as a major product and ω-hydroxy-lauric acid as a minor product. Steady-state kinetic analysis of lauric acid hydroxylation yielded a kcat value of 0.038 ± 0.002 min-1 and a Km value of 10 ± 2 µM. In addition, capric acid hydroxylation of CYP6A8 yielded kinetic parameters with a kcat value of 0.135 ± 0.007 min-1 and a Km value of 21 ± 4 µM. Because of the importance of various lipids as carbon sources, the metabolic analysis of fatty acids using CYP6A8 in this study can provide an understanding of the biochemical roles of P450 enzymes in many insects, including Drosophila melanogaster.

Hyaluronic acid and proteoglycan link protein 1 suppresses platelet-derived growth factor-BB-induced proliferation, migration, and phenotypic switching of vascular smooth muscle cells

  • Dan Zhou;Hae Chan Ha;Goowon Yang;Ji Min Jang;Bo Kyung Park;Bo Kyung Park;In Chul Shin;Dae Kyong Kim
    • BMB Reports
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    • v.56 no.8
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    • pp.445-450
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    • 2023
  • The development of atherosclerotic cardiovascular disease is associated with the phenotypic switching of vascular smooth muscle cells (SMCs) from a contractile to a synthetic state, leading to cell migration and proliferation. Platelet-derived growth factor-BB (PDGF-BB) modulates this de-differentiation by initiating a number of biological processes. In this study, we show that gene expression of hyaluronic acid (HA) and proteoglycan link protein 1 (HAPLN1) was upregulated during differentiation of human aortic SMCs (HASMCs) into a contractile state, but downregulated upon during PDGF-BB-induced dedifferentiation. This is the first study showing that the treatment of HASMCs with full-length recombinant human HAPLN1 (rhHAPLN1) significantly reversed PDGF-BB-induced decrease in the protein levels of contractile markers (SM22α, α-SMA, calponin, and SM-MHC), and inhibited the proliferation and migration of HASMCs induced by PDGF-BB. Furthermore, our results show that rhHAPLN1 significantly inhibited the phosphorylation of FAK, AKT, STAT3, p38 MAPK and Raf mediated by the binding of PDGF-BB to PDGFRβ. Together, these results indicated that rhHAPLN1 can suppress the PDGF-BB-stimulated phenotypic switching and subsequent de-differentiation of HASMCs, highlighting its potential as a novel therapeutic target for atherosclerosis and other vascular diseases.

Interaction of Proline with Cu+ and Cu2+ Ions in the Gas Phase (기체상에서 Cu+ 및 Cu2+ 이온과 proline의 상호작용)

  • Lee, Gab-Yong
    • Journal of the Korean Chemical Society
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    • v.53 no.3
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    • pp.257-265
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    • 2009
  • The structures and metal affinities of the binding configurations of $Cu^{+}$ and $Cu^{2+}$ to proline have been investigated using the hybrid three-parameter Density Functional Theory(DFT/B3LYP). We found that the metal-proline bonding and the energy ordering of several conformers were very different in $Cu^{+}$-proline and $Cu^{2+}$-proline. For $Cu^{+}$-proline, the ground state structure was found to have a bidentated coordination in which $Cu^{+}$ was coordinated to the carbonyl oxygen and imino group nitrogen of neutral proline. On the contrary, the ground state structure of $Cu^{2+}$-proline involves chelation between the two oxygens of the carboxylate group in a zwitterionic proline. The metal ion affinity of proline of the most stable $Cu^{+}$-proline complex was calculated as 76.0 kcal/mol at 6-311++G(d,p) level, whereas the $Cu^{2+}$ ion affinity of proline was calculated as 258.5 kcal/mol.

Biochemical Characterization of the Dual Positional Specific Maize Lipoxygenase and the Dependence of Lagging and Initial Burst Phenomenon on pH, Substrate, and Detergent during Pre-steady State Kinetics

  • Cho, Kyoung-Won;Jang, Sung-Kuk;Huon, Thavrak;Park, Sang-Wook;Han, Ok-Soo
    • BMB Reports
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    • v.40 no.1
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    • pp.100-106
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    • 2007
  • The wound-inducible lipoxygenase obtained from maize is one of the nontraditional lipoxygenases that possess dual positional specificity. In this paper, we provide our results on the determination and comparison of the kinetic constants of the maize lipoxygenase, with or without detergents in the steady state, and characterization of the dependence of the kinetic lag phase or initial burst, on pH, substrate, and detergent in the pre-steady state of the lipoxygenase reaction. The oxidation of linoleic acid showed a typical lag phase in the pre-steady state of the lipoxygenase reaction at pH 7.5 in the presence of 0.25% Tween-20 detergent. The reciprocal correlation between the induction period and the enzyme level indicated that this lag phenomenon was attributable to the slow oxidative activation of Fe (II) to Fe (III) at the active site of the enzyme as observed in other lipoxygenase reactions. Contrary to the lagging phenomenon observed at pH 7.5 in the presence of Tween-20, a unique initial burst was observed at pH 6.2 in the absence of detergents. To our knowledge, the initial burst in the oxidation of linoleic acid at pH 6.2 is the first observation in the lipoxygenase reaction. Kinetic constants (Km and kcat values) were largely dependent on the presence of detergent. An inverse correlation of the initial burst period with enzyme levels and interpretations on kinetic constants suggested that the observed initial burst in the oxidation of linoleic acid could be due to the availability of free fatty acids as substrates for binding with the lipoxygenase enzyme.

Electrochemical Characterization of Multilayered CdTe/PSS Films Prepared by Electrostatic Self-assembly Method

  • Rabbani, Mohammad Mahbub;Yeum, Jeong Hyun;Kim, Jungsoo;Nam, Dae-Geun;Oh, Weontae
    • Transactions on Electrical and Electronic Materials
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    • v.15 no.5
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    • pp.257-261
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    • 2014
  • Multilayered CdTe/PSS films were prepared by the electrostatic self-assembly method in an aqueous medium. Positively-charged cadmium telluride (CdTe) nanoparticles and anionic polyelectrolyte, poly (sodium 4-styrene sulfonate) (PSS) were assembled alternately in order to build up a multilayered film structure. A linear proportion of absorbance to the number of bilayers suggests that an equal amount of CdTe was adsorbed after each dipping cycle, which resulted in the buildup of a homogenous film. The binding energies of elements (Cd and Te) in multilayered CdTe/PSS film shifted from those of the CdTe nanoparticles in the pure state. This result indicates that the interfacial electron densities were redistributed by the strong electrostatic interaction between the oppositely-charged CdTe and PSS. Electrochemical properties of the multilayered CdTe/PSS films were studied in detail by cyclic voltammetry (CV).

Interfacial Natures and Controlling Morphology of Co Oxide Nanocrystal Structures by Adding Spectator Ni Ions

  • Gwag, Jin-Seog;Sohn, Young-Ku
    • Bulletin of the Korean Chemical Society
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    • v.33 no.2
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    • pp.505-510
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    • 2012
  • Cobalt oxide nanostructure materials have been prepared by adding several concentrations of spectator Ni ions in solution, and analyzed by electron microscopy, X-day diffraction, calorimetry/thermogravimetric analysis, UV-vis absorption, Raman, and X-ray photoelectron spectroscopy. The electron microscopy results show that the morphology of the nanostructures is dramatically altered by changing the concentration of spectator ions. The bulk XRD patterns of $350^{\circ}C$-annealed samples indicate that the structure of the cobalt oxide is all of cubic Fd-3m $Co_3O_4$, and show that the major XRD peaks shift slightly with the concentration of Ni ions. In Raman spectroscopy, we can confirm the XRD data through a more obvious change in peak position, broadness, and intensity. For the un-sputtered samples in the XPS measurement process, the XPS peaks of Co 2p and O 1s for the samples prepared without Ni ions exhibit higher binding energies than those for the sample prepared with Ni ions. Upon $Ar^+$ ion sputtering, we found $Co_3O_4$ reduces to CoO, on the basis of XPS data. Our study could be further applied to controlling morphology and surface oxidation state.