• Title/Summary/Keyword: Dimeric interaction

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Mode of Action on EcoRI Restriction Endonuclease: EcoRI and EcoRI Variant N199H have Active Monomeric Forms

  • Kim, Jae-Jong;Koh, Suk-Hoon;Kim, Joong-Su;Lee, Dae-Sil
    • BMB Reports
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    • v.31 no.2
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    • pp.149-155
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    • 1998
  • The N199H variant of the EcoRI endonuclease has about twice the catalytic activity of the wild-type. A comparison of their biochemical characteristics, using synthetic oligonucleotides 5'-dAAAACTTAAGAAAAAAAAAAA-3' (KA) and 5'-dTTTTTGAATTCTTTTTTTTTT-3' (KT), helps to define the cleavage reaction pathway of these enzymes. Both EcoRI and EcoRI variant N199H were found to cleave singlestranded KA or KT about three times faster than the double-stranded forms, although the KT oligonucleotide was more susceptible. Using the ssDNA substrate in kinetic analyses, lower $K_m$ values were obtained for the N199H variant than for the wild-type at low (50 mM), as well as high (200 mM), sodium chloride concentrations. This difference between the endonucleases is attributed to a grealter accessibility for tbe substrate by the variant, and also a higher affinity for the DNA backbone. It also appears that the relative activities of the two enzymes, particularly at high ionic strength, are proportional to their populations in the monomeric enzyme form. That is, according to gel filtration data, half of the N199H molecules exist as monomers in 200 mM NaCl, whereas those of the wild-type are mainly dimeric. Consequently, the Asp199 residue of the EcoRI endonuclease may be implicated in the protein-protein interaction leading to dimerization, as well as in coupling to DNA substrates. In summary, it is proposed that active monomeric endonuclease molecules, derived from the dimeric enzyme, recognize and form a complex with a single stranded form of the DNA substrate, which then undergoes nucleophilic substitution and cleavage.

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Copper(II) Coordination Polymers Assembled from 2-[(Pyridin-3-ylmethyl)amino]ethanol: Structure and Magnetism

  • Han, Jeong-Hyeong;Shin, Jong-Won;Min, Kil-Sik
    • Bulletin of the Korean Chemical Society
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    • v.30 no.5
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    • pp.1113-1117
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    • 2009
  • The one-dimensional coordination polymers, $[Cu^{II}(L)(NO_3)_2]_n$ (1) and {$[Cu^{II}(L)(NO_3)]{\cdot}2H_2O}_{2n} (2), were synthesized from $Cu(NO_3)_2{\cdot}3H_2O$ and 2-[(pyridin-3-ylmethyl)amino]ethanol (L, PMAE) in methanol by controlling the molar ratio of copper(II) salt. Copper(II) ion in 1 has one pyridine group of PMAE whose an aminoethanol group coordinates adjacent copper(II) ion. As the pyridine group is bonded to neighboring copper(II) ion, 1 becomes a one-dimensional chain. Contrary to 1, the structure of 2 shows that the oxygen atom of ethoxide group is bridged between two copper(II) ions, which forms a dinuclear complex. Additionally, the pyridine group of PMAE included one dinuclear unit is coordinated to the other dimeric one each other, which leads to a one-dimensional polymer. Due to the structural differences, 1 exhibits weak antiferromagnetic interaction, while 2 shows strong antiferromagnetic interaction. Due to direct spin exchange via oxygen of PMAE 2 has a much strong spin coupling than 1.

Template Synthesis, Crystal Structure, and Magnetic Properties of a Dinuclear Copper(II) Complex with Cooperative Hydrogen Bonding

  • Kang, Shin-Geol;Nam, Kwang-Hee;Min, Kil-Sik;Lee, Uk
    • Bulletin of the Korean Chemical Society
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    • v.32 no.3
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    • pp.1037-1040
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    • 2011
  • The dinuclear complex 1 with cooperative hydrogen bonds can be prepared by the metal-directed reaction of Eq. (2). This work shows that the coordinated hydroxyl group trans to the secondary amino group is deprotonated more readily than that trans to the tertiary amino group and acts as the hydrogen-bond accepter. The lattice water molecules in 1 act as bridges between the two mononuclear units through hydrogen bonds. The complex is quite stable as the dimeric form even in various polar solvents. The complex exhibits a weak antiferromagnetic interaction between the metal ions in spite of relatively long Cu$\cdots$Cu distance. This strongly supports the suggestion that the antiferromagnetic behavior is closely related to the cooperative hydrogen bonds.

$^{13}C\;and\;^{51}V $ Nuclear Magnetic Resonance Studies of Vanadium (Ⅴ)-$\alpha$-Hydroxycarboxylate Complexes

  • Man-Ho Lee;In-Whan Kim
    • Bulletin of the Korean Chemical Society
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    • v.14 no.5
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    • pp.557-561
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    • 1993
  • The interaction of vanadium(V) with various a-hydroxycarboxylate ligands in aqueous solution at pH 3.2 have been studied by $^{51}V$ and $^{13}C$ NMR spectroscopies. From the results it is supposed that vanadates mainly form the octahedral complexes with lactate, 2-hydroxybutyrate, glycerate, and malate. While, vanadates form the trigonal-bipyramidal complexes with glycolate, tartarate, and 2-hydroxy-3-methylbutyrate, and tetrahedral complexes with pyruvate(diol), 2-hydroxyisobutyrate, and 2-hydroxy-3-methylbutyrate. The bipyramidal products are formed as monomeric compounds. The octahedral products are formed as dimeric compounds with no evidence for a significant proportion of the monomeric derivatives. The complexes are mainly formed through the coordination at the carboxylate and the 2-hydroxyl groups of the ligands.

Homo- or Hetero-Dimerization of Muscarinic Receptor Subtypes is Not Mediated by Direct Protein-Protein Interaction Through Intracellular and Extracellular Regions

  • Kang, Yun-Kyung;Yoon, Tae-Sook;Lee, Kyung-Lim;Kim, Hwa-Jung
    • Archives of Pharmacal Research
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    • v.26 no.10
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    • pp.846-854
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    • 2003
  • The oligomerization of G-proteincoupled receptors (GPCRs) has been shown to occur by various mechanisms, such as via disulfide covalent linkages, non covalent (ionic, hydrophobic) interactions of the N-terminal, and/or transmembrane and/or intracellular domains. Interactions between GPCRs could involve an association between identical proteins (homomers) or non-identical proteins (heteromers), or between two monomers (to form dimers) or multiple monomers (to form oligomers). It is believed that muscarinic receptors may also be arranged into dimeric or oigomeric complexes, but no systematic experimental evidence exists concerning the direct physical interaction between receptor proteins as its mechanism. We undertook this study to determine whether muscarinic receptors form homomers or a heteromers by direct protein-protein interaction within the same or within different subtypes using a yeast two-hybrid system. Intracellular loops (i1, i2 and i3) and the C-terminal cytoplasmic tails (C) of human muscarinic (Hm) receptor subtypes, Hm1, Hm2 and Hm3, were cloned into the vectors (pB42AD and pLexA) of a two-hybrid system and examined for heteromeric or homodimeric interactions between the cytoplasmic domains. No physical interaction was observed between the intracellular domains of any of the Hm/Hm receptor sets tested. The results of our study suggest that the Hm1, Hm2 and Hm3 receptors do not form dimers or oligomers by interacting directly through either the hydrophilic intracellular domains or the C-terminal tail domains. To further investigate extracellular domain interactions, the N-terminus (N) and extracellular loops (o1 and o2) were also cloned into the two-hybrid vectors. Interactions of Hm2N with Hm2N, Hm2o1, Hm2o2, Hm3N, Hm3o1 or Hm3o2 were examined. The N-terminal domain of Hm2 was found to have no direct interaction with any extracellular domain. From our results, we excluded the possibility of a direct interaction between the muscarinic receptor subtypes (Hm1, Hm2 and Hm3) as a mechanism for homo- or hetero-meric dimerization/oligomerization. On the other hand, it remains a possibility that interaction may occur indirectly or require proper conformation or subunit formation or hydrophobic region involvement.

Structural, Electrochemical, DNA Binding and Cleavage Properties of Nickel(II) Complex [Ni(H2biim)2(H2O)2]2+ of 2,2'-Biimidazole

  • Jayamani, Arumugam;Thamilarasan, Vijayan;Ganesan, Venketasan;Sengottuvelan, Nallathambi
    • Bulletin of the Korean Chemical Society
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    • v.34 no.12
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    • pp.3695-3702
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    • 2013
  • A nickel(II) complex $[Ni(H_2biim)_2(H_2O)_2](ClO_4)_2{\cdot}H_2O$ (1) of biimidazole ligand has been synthesized and characterized (Where $H_2biim$ = 2,2'-biimidazole). The single crystal X-ray diffraction of the complex shows a dimeric structure with six coordinated psudo-octahedral geometry. The cyclic voltammograms of complex exhibited one quasireversible reduction wave ($E_{pc}=-0.61V$) and an irreversible oxidation wave ($E_{pa}=1.28V$) in DMF solution. The interaction of the complex with Calf-Thymus DNA (CT-DNA) has been investigated by absorption and fluorescence spectroscopy. The complex is an avid DNA binder with a binding constant value of $1.03{\times}10^5M^{-1}$. The results suggest that the nickel(II) complex bind to CT-DNA via intercalative mode and can quench the fluorescence intensity of EB bind to CT-DNA with $K_{app}$ value of $3.2{\times}10^5M^{-1}$. The complex also shown efficient oxidative cleavage of supercoiled pBR322 DNA in the presence of hydrogen peroxide as oxidizing agent. The DNA cleavage by complex in presence of quenchers; viz. DMSO, KI, $NaN_3$ and EDTA reveals that hydroxyl radical or singlet oxygen mechanism is involved. The complex showed invitro antimicrobial activity against four bacteria and two fungi. The antimicrobial activity was nearer to that of standard drugs and greater than that of the free ligand.

Oligomeric Structures Determine the Biochemical Characteristics of Human Nucleoside Diphosphate Kinases

  • Kim, Sun-Young;Song, Eun-Joo;Chang, Keun-Hye;Kim, Eun-Hee;Chae, Suhn-Kee;Lee, Han-Soo;Lee, Kong-Joo
    • BMB Reports
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    • v.34 no.4
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    • pp.355-364
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    • 2001
  • Major human Nucleoside diphosphate kinases (NDPKs) exist as hetero-oligomers, consisting of NDPK-A and NDPK-B, rather than homo-oligomer. To investigate their biological function depending on the oligomeric structure in vivo, we characterized the biochemical properties of cellular NDPK. Cellular NDPKs, which are made up of a unique combination of isoforms, were purified from human erythrocyte and placenta. We found that cellular NDPK and recombinant isoforms NDPKs have their own distinct biochemical properties in autophosphorylation, stability toward heat or urea, and DNA binding. Cellular NDPK was found to have unique characteristics rather than the expected additive properties of recombinant isoforms. The mutations in the dimeric interface of NDPK-B (R34G, N69H or K135L) caused defective DNA binding and simultaneously reduced the enzymatic stability These results suggest that the oligomeric interaction could play a major role in the stability of catalytic domain and might be related to the regulation of various cellular functions of NDPK.

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Semi-Empirical MO Calculations and Infrared Spectroscopy of Hydrogen-Bonding in Alkyl Alcohols (알킬알코올의 수소결합에 대한 적외선분광법 및 반경험적분자궤도함수의 계산)

  • Jong Taik Kim;Doo Seon Park
    • Journal of the Korean Chemical Society
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    • v.31 no.1
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    • pp.3-13
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    • 1987
  • Infrared absorption spectra of alkyl alcohols in the OH stretching region were obtained from varying the concentrations of alcohols in $CCl_4$. The OH stretching bands were broadened and shifted to lower frequencies due to the hydrogen-bond formation. Three bands were obtained from the breakdown of these bands by the simplex method. Each band was assigned to various types of hydrogen-bonded OH groups. The electronic structures and interaction energies of dimeric and trimeric alcohols were calculated by semi-empirical MO(CNDO/2, INDO) methods. These results were in good agreement with those of deconvoluted ir spectra. The EDA(electron donor-acceptor) effect of alkyl group on hydrogen-bond formation was in the decreasing order of butyl > propyl > ethyl > methyl group. On the other hand, the experimental results were in the order : propyl > ethyl > butyl > methyl group. This seemed to be ascribed to the bulkiness of butyl alcohol.

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Regulation of Gene Expression and 3-Dimensional Structure of DNA (유전자 발현 조절과 DNA 3차원적 구조와의 관계)

  • 김병동
    • Proceedings of the Botanical Society of Korea Conference
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    • 1987.07a
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    • pp.149-155
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    • 1987
  • Growth and development of a higher plant, or any living organism for that matter, could be defined as an orderly expression of the genome in time and space in close interaction with the environment. During differentiation and development of a tissue or organ a group of genes must be selectively turned on or turned off mainly by trans-acting regulators. In this general concept of regulation of regulation of gene expression, a DNA molecule is recognized at a specific nucleotide sequence by DNA-binding factors. Molecular biology of the regulatory factors such as hormones, and their receptors, target DNA sequences and DNA-binding proteins are well advanced. What is not clearly understood is the molecular basis of the interactions between DNA and binding factors, expecially of the usages of the dyad symmetry of the target DNA sequences and the dimeric nature of the DNA-binding proteins. A unique 3-dimensional structure of DNA has been proposed that may play an important role in the orderly expression of the gene. A foldback intercoil (FBI) DNA configuration which was originally found by electron microscopy among mtDNA molecules from pearl millet has some unique features. The FBI configuration of DNA is believed to be formed when a flexible double helix folds back and interwines in the widened major grooves resulting in a four stranded, intercoil DNA whose thickness is the same as that of double stranded DNA. More recently, the FBI structure of DNA has been also induced in vitro by a novel enzyme which was purified from pearl millet mitochondria. It has been proposed that the FBI DNA could be utillized in intramolecular recombination which leads to inversion or deletion, and in intermolecular recombination which can lead to either site-specific recombination, genetic recombination via single strand invasion, or cross strand recombination. The structure and function of DNA in 3-dimensional aspect is emphasized for better understanding orderly expression of genes during growth and development.

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