• Title/Summary/Keyword: Binding Interface

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Mutation of a Transposed Amino Acid Triplet Repeat Enhances Coupling of m1 Muscarinic Receptor to Activation of Phospholipase C

  • Lee, Seok-Yong;Cho, Tai-Soon
    • Proceedings of the Korean Society of Applied Pharmacology
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    • 1996.04a
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    • pp.206-206
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    • 1996
  • The C-terminus ends of the second putative transmembrane domains of both m1 and m2 muscarinic receptors contain a triplet of amino acid residues consisting of leucine (L), tyrosine (Y) and threonine (T), This triplet is repeated as LYT-LYT in m2 receptors at the interface between the second transmembrane domain and the first extracellular loop. Interestingly, however, it is repeated in a transposed fashion (LYT-TYL) in the sequence of m1 receptors. In this work we employed site-directed mutagenesis to investigate the possible significance of this unique sequence diversity for determining the distinct differential drug-receptor interaction and cellular function at m1 muscarinic receptor. Mutation of the LYTTYL sequence of m1 receptors to the corresponding m2 receptor LYTLYT sequence, however, did not result in a significant change in the binding affinity of the agonist carbachol or in the affinity of the majority of a series of receptor antagonists which are able to discriminate between wild-type m1 and m2 receptors. Surprisingly, the LYTLYT ml receptor mutant demonstrated markedly enhanced coupling to activation of phospholipase C without a change in its coupling to increased cyclic AMP formation. There was also an enhanced receptor sensitivity in transducing elevation of intracellular Ca$\^$2+/. These changes were not due to alterations in the rate of receptor. desensitization or sequestration, On the other hand, the reverse LYTLYT-LYTTYL mutation in the m2 receptor did not alter its coupling to inhibition of adenylate cyclase, but slightly enhanced its coupling to stimulation of PI hydrolysis, Our data suggest that the LYTTYL/LYTLYT sequence difference between ml and n12 muscarinic receptors is not involved in determining receptor pharmacology. On the other hand, while these differences might play a role in the modulation of muscarinic receptor coupling to PI hydrolysis, they are not important for specifying coupling of various subtypes of muscarinic receptors to different cellular signaling pathways.

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Study on the micellization of cetyltrimethyl ammonium bromide in 4-biphenyl acetate solution (4-biphenyl acetate 수용액에서 Cetyltrimethyl Ammonium Bromide의 미셀화에 관한 연구)

  • Oh, Jung Hee
    • Analytical Science and Technology
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    • v.8 no.2
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    • pp.107-116
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    • 1995
  • The critical micelle concentration(CMC) of CTAB was determined with changes in absorbance at 202nm band of 4-biphenyl acetate($BPA^-$). With $BPA^-$ as a probe, the effect of temperature on CMC of CTAB has been observed between $30^{\circ}C{\sim}70^{\circ}C$. In this range of temperature the values of CMC are $1.18{\times}10^{-4}{\sim}2.02{\times}10^{-4}M$. The free energy(${\Delta}G^{\circ}m$) and enthalpy(${\Delta}H^{\circ}m$)for the micellization of CTAB was negative and the entropy(${\Delta}S^{\circ}m$) was a large positive value. The micellization of CTAB is considered as a spontaneous process and to involve a phase transition. The orientational binding of 4-biphenyl acetate anion to the CTAB micelle interface has been studied with $300MHz\;H^1-NMR$ data. The change in chemical shift of proton in CTAB as well as those of the protons in $BPA^-$ have been investigated by increasing the mole fraction of the anion in the mixed solutions. The changes in chemical shift with increasing mole fraction of anion($BPA^-$) indicate the formation of mixed micelle between CTAB and $BPA^-$. The changes in chemical shifts of methylene protons in CTAB, demonstrate the penetration of $BPA^-$ into the palisade layer of the CTAB micelle.

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A Design Method for Dynamic Selection of SOA Services (SOA 서비스의 동적 선택 설계 기법)

  • Bae, Jeong-Seop;La, Hyun-Jung;Kim, Soo-Dong
    • Journal of KIISE:Software and Applications
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    • v.35 no.2
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    • pp.91-104
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    • 2008
  • Service-Oriented Computing (SOC) is the development method that published services are selected and composed at runtime to deliver the expected functionality to service clients. SOC should get maximum benefits not only supporting business agility but also reducing the development time. Services are selected and composed at runtime to improve the benefits. However, current programming language, SOC platforms, business process modeling language, and tools support either manual selection or static binding of published services. There is a limitation on reconfiguring and redeploying the business process to deliver the expected services to each client. Therefore, dynamic selection is needed for composing appropriate services to service clients in a quick and flexible manner. In this paper, we propose Dynamic Selection Handler (DSH) on ESB. we present a design method of Dynamic Selection Handler which consists of four components; Invocation Listener, Service Selector, Service Binder and Interface Transformer. We apply appropriate design patterns for each component to maximize reusability of components. Finally, we describe a case study that shows the feasibility of DSH on ESB.

Microstructural and Wear Properties of WC-based and Cr3C2-based Cermet Coating Materials Manufactured with High Velocity Oxygen Fuel Process (고속 화염 용사 공정으로 제조된 WC계 및 Cr3C2계 Cermet 코팅 소재의 미세조직 및 마모 특성)

  • Kang, Yeon-Ji;Ham, Gi-Su;Kim, Hyung-Jun;Yoon, Sang-Hoon;Lee, Kee-Ahn
    • Journal of Powder Materials
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    • v.25 no.5
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    • pp.408-414
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    • 2018
  • This study investigates the microstructure and wear properties of cermet (ceramic + metal) coating materials manufactured using high velocity oxygen fuel (HVOF) process. Three types of HVOF coating layers are formed by depositing WC-12Co, WC-20Cr-7Ni, and Cr3C2-20NiCr (wt.%) powders on S45C steel substrate. The porosities of the coating layers are $1{\pm}0.5%$ for all three specimens. Microstructural analysis confirms the formation of second carbide phases of $W_2C$, $Co_6W_6C$, and $Cr_7C_3$ owing to decarburizing of WC phases on WC-based coating layers. In the case of WC-12Co coating, which has a high ratio of $W_2C$ phase with high brittleness, the interface property between the carbide and the metal binder slightly decreases. In the $Cr_3C_2-20CrNi$ coating layer, decarburizing almost does not occur, but fine cavities exist between the splats. The wear loss occurs in the descending order of $Cr_3C_2-20NiCr$, WC-12Co, and WC-20Cr-7Ni, where WC-20Cr-7Ni achieves the highest wear resistance property. It can be inferred that the ratio of the carbide and the binding properties between carbide-binder and binder-binder in a cermet coating material manufactured with HVOF as the primary factors determine the wear properties of the cermet coating material.

Prediction of Protein-Protein Interaction Sites Based on 3D Surface Patches Using SVM (SVM 모델을 이용한 3차원 패치 기반 단백질 상호작용 사이트 예측기법)

  • Park, Sung-Hee;Hansen, Bjorn
    • The KIPS Transactions:PartD
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    • v.19D no.1
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    • pp.21-28
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    • 2012
  • Predication of protein interaction sites for monomer structures can reduce the search space for protein docking and has been regarded as very significant for predicting unknown functions of proteins from their interacting proteins whose functions are known. In the other hand, the prediction of interaction sites has been limited in crystallizing weakly interacting complexes which are transient and do not form the complexes stable enough for obtaining experimental structures by crystallization or even NMR for the most important protein-protein interactions. This work reports the calculation of 3D surface patches of complex structures and their properties and a machine learning approach to build a predictive model for the 3D surface patches in interaction and non-interaction sites using support vector machine. To overcome classification problems for class imbalanced data, we employed an under-sampling technique. 9 properties of the patches were calculated from amino acid compositions and secondary structure elements. With 10 fold cross validation, the predictive model built from SVM achieved an accuracy of 92.7% for classification of 3D patches in interaction and non-interaction sites from 147 complexes.

Molecular Structures and Catalytic Mechanism of Bacterial Lipases. (세균성 리파제의 분자구조와 작용기작)

  • 김형권
    • Microbiology and Biotechnology Letters
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    • v.31 no.4
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    • pp.311-321
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    • 2003
  • Bacteria produce lipases, which can catalyze both the hydrolysis and the synthesis of long chain triglycerides. These reactions usually proceed with high regioselectivity and enantioselectivity, and, therefore, lipases have become very important biocatalysts used in organic chemistry. 3D lipase structures were solved from several bacterial lipases. They have an $\alpha/\beta$ hydrolase fold and a catalytic triad consisting of a nucleophilic serine, and an aspartate or glutamate residue that is hydrogen bonded to a histindine. Active sites are covered with $\alpha$-helical lid structure, of which movement is involved in the enzyme's activation at oil/water interface. Four substrate binding pockets were identified for triglycerides: an oxyanion hole and three pockets accommodating the fatty acids bound at positions sn-1, sn-2, and sn-3. These pockets determine the enantiopreference of a lipase. The understanding of structure-function relationships as well as the development of molecular evolution techniques will enable researchers to tailor new lipases for biotechnological applications.

Electrophoretic Mobility to Monitor Protein-Surfacant Interactions

  • Hong, Soon-Taek
    • Preventive Nutrition and Food Science
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    • v.3 no.2
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    • pp.143-151
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    • 1998
  • Protein -surfactant interactions have been investigate by measuring ζ-potential of $\beta$-lactoglobulin-coated emulsion droplets and $\beta$-lactoglobulin in solution in the rpesenceof surfactant, with particular emphasis on the effect of protein heat treatment(7$0^{\circ}C$, 30min). When ionic surfactant (SDS or DATEM) is added to the protein solution, the ζ-potential of the mixture is found to increase with increasing surfactant concentration, indicating surfactant binding to the protein molecules. For heat-denatured protein,it has been observed that the ζ-potential tends to be lower than that of the native protein. The effect of surfactant on emulsions is rather complicated .With SDS, small amounts of surfactant addition induce a sharp increase in zeta potential arising from the specific interaction of surfactant with protein. With further surfacant addition, there is a gradual reductio in the ζ-potential, presumably caused by the displacement of adsorped protein (and protein-surfactant complex) from the emulsion droplet surfac by the excess of SDS molecules. At even higher surfactant concentrations, the measured zeta potential appears to increase slightly, possibly due to the formation of a surfactant measured zeta potential appears to increase slightly, possibly due to the formation of surfactant micellar structure at the oil droplet surface. This behaviour contrastswith the results of the corresponding systems containing the anionic emulsifier DATEM, in which the ζ-potential of the system is found to increase continuously with R, particularly at very low surfactant concentration. Overall, such behaviour is consisten with a combination of complexation and competitive displacement between surfactant and protein occurring at the oil-water interface. In addition, it has also been found that above the CMC, there is a time-dependent increase in the negative ζ-potential of emulsion droplets in solutions of SDS, possibly due to the solublization of oil droplets into surfactant micelles in the aqueous bulk phase.

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Effect of Plasma Treatment Times on the Adhesion of Cu/Ni Thin Film to Polyimide (폴리이미드와 Cu/Ni층과의 계면결합력에 미치는 플라즈마 처리 시간 효과)

  • Woo, Tae-Gyu;Park, Il-Song;Jung, Kwang-Hee;Jeon, Woo-Yong;Seol, Kyeong-Won
    • Korean Journal of Metals and Materials
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    • v.49 no.8
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    • pp.657-663
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    • 2011
  • This study represents the results of the peel strength and surface morphology according to the preprocessing times of polyimide (PI) in a Cu/Ni/PI structure flexible copper clad laminate production process based on the polyimide. Field emission scanning electron microscopy, X-ray diffraction, and X-ray photoelectron spectroscopy were used to analyze the surface morphology, crystal structure, and interface binding structure of sputtered Ni, Cu, and electrodeposited copper foil layers. The surface roughness of Ni, Cu deposition layers and the crystal structure of electrodeposited Cu layers were varied according to the preprocessing times. In the RF plasma times that were varied by 100-600 seconds in a preprocessing process, the preprocessing applied by about 300-400 seconds showed a homogeneous surface morphology in the metal layers and that also represented high peel strength for the polyimide. Considering the effect of peel strength on plastic deformation, preprocessing times can reasonably be at about 400 seconds.

Visualization of Geometric Features in the Contact Region of Proteins (단백질 접촉 영역의 기하학적 특성 가시화)

  • Kim, Ku-Jin
    • KIPS Transactions on Software and Data Engineering
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    • v.8 no.10
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    • pp.421-426
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    • 2019
  • In this paper, we propose a method to visualize the geometric features of the contact region between proteins in a protein complex. When proteins or ligands are represented as curved surfaces with irregularities, the property that the two surfaces contact each other without intersections is called shape compatibility. Protein-Protein or Protein-Ligand docking researches have shown that shape complementarity, chemical properties, and entropy play an important role in finding contact regions. Usually, after finding a region with high shape complementarity, we can predict the contact region by using residual polarity and hydrophobicity of amino acids belonging to this region. In the research for predicting the contact region, it is necessary to investigate the geometrical features of the contact region in known protein complexes. For this purpose, it is essential to visualize the geometric features of the molecular surface. In this paper, we propose a method to find the contact region, and visualize the geometric features of it as normal vectors and mean curvatures of the protein complex.

Cadmium-Substituted Concanavalin A and Its Trimeric Complexation

  • Park, Yeo Reum;Kim, Da Som;Lee, Dong-Heon;Kang, Hyun Goo;Park, Jung Hee;Lee, Seung Jae
    • Journal of Microbiology and Biotechnology
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    • v.28 no.12
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    • pp.2106-2112
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    • 2018
  • Concanavalin A (ConA) interacts with carbohydrates as a lectin, and recent reports proposed its application for detecting a diversity of viruses and pathogens. Structural studies have detailed the interaction between ConA and carbohydrates and the metal coordination environment with manganese and calcium ions (Mn-Ca-ConA). In this study, ConA was crystallized with a cadmium-containing precipitant, and the refined structure indicates that $Mn^{2+}$ was replaced by $Cd^{2+}$ (Cd-Ca-ConA). The structural comparison with ConA demonstrates that the metal-coordinated residues of Cd-Ca-ConA, that is Glu8, Asp10, Asn14, Asp19, and His24, do not have conformational shifts, but residues for sugar binding, including Arg228, Tyr100, and Leu99, reorient their side chains, slightly. Previous studies demonstrated that excess cadmium ions can coordinate with other residues, including Glu87 and Glu183, which were not coordinated with $Cd^{2+}$ in this study. The trimeric ConA in this study coordinated $Cd^{2+}$ with other residues, including Asp80 and Asp82, for complex generation. The monomer does not have specific interaction near interface regions with the other monomer, but secondary cadmium coordinated with two aspartates (Asp80 and Asp82) from monomer 1 and one aspartate (Asp16) from monomer 2. This study demonstrated that complex generation was induced via coordination with secondary $Cd^{2+}$ and showed the application potential regarding the design of complex formation for specific interactions with target saccharides.