• 제목/요약/키워드: Binding property

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Generation and characterization of calmodulin-DHFR sandwich fusion protein

  • Han, Chang Hoon
    • Korean Journal of Veterinary Research
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    • v.48 no.3
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    • pp.243-250
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    • 2008
  • A calmodulin-dihydrofolate reductase (DHFR) sandwich fusion protein was generated by insertion of calmodulin into the $\beta$-bulge region of DHFR to observe the effects of structurally constraining the calmodulin structure. The calcium binding properties of the sandwich protein were almost identical to calmodulin. Similar to calmodulin ($10.7 {\mu}M$), the sandwich protein bound four equivalents of calcium, with half saturation ($K_{0.5}$) observed at a [$Ca^{2+}$] of $8{\mu}M$. However, nicotinamide adenine dinucleotide (NAD) kinase activation property of the sandwich protein was lower than that of calmodulin. The sandwich protein activated NAD kinase, but to only half of the level obtained with calmodulin. The K 0.5 for both calmodulin and the sandwich protein were approximately the same (1-2 nM). Methylation analyses of the sandwich protein show that insertion of calmodulin into DHFR results in a large decrease in methylation. The $V_{max}$ observed with the sandwich protein (95 nmole/min/ml) was only 22% of the value observed with calmodulin (436 nmol/min/ml) in the presence of calcium. Addition of trimethoprim to the reaction significantly inhibited the observed methylation rate. Overall, the data suggest that the insertion of calmodulin into the DHFR structure has little effect on calcium binding by the individual lobes of calmodulin, but may constrain the lobes in a manner that results in altered interaction with the calmodulin-dependent proteins, and severely perturbed the methyltransferase recognition site.

Mutational Analysis of Two Conserved Active Site Tyrosine Residues in Matrilysin

  • Jaeho Cha
    • Journal of Life Science
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    • v.9 no.2
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    • pp.44-48
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    • 1999
  • The ionization of tyrosine residue is known to be involved in the stabilization of transition-state in catalysis of astacin based upon the astacin-transition state analog structure. Two tyrosine residues, Tyr-216 and Tyr-219, are conserved in all MMPs related with astacin family, We replaced Tyr-216 and Tyr-219 into phenylalanine, respectively and the zinc binding properties, kinetic parameters, and pH dependence of each mutant are determined in order to examine the role of tyrosine residue in matrilysin catalysis. Both mutants contain two zinc atoms per mol of enzyme, indicating that either tyrosime does not affect the zinc binding property of the enzyme. Y216F and Y219F mutants are highly active and the kcat/Km values are only decreased 1.1-1.5-fold compared to the wild-type enzyme. The decrease in the activity of the mutants is essentially due to the increase in Km value. The pH dependencies of the kcat/Km values for both mutants are similar to the corresponding dependencies obtained with the wild type enzyme. The pKa values at the alkaline side of both mutants are not changed. These kinetic and pH dependence results indicate that the ionization of active site tyrosine residue of matrilysin is not reflected in the kinetics of peptide hydrolysin as catalyzed by astacin.

A Study on the Binding Force of Drawbead in the Sheet Metal Forming Process through the finite element and experimental analysis (해석과 실험을 통한 박판성형공정에서의 드로오비드의 구속력에 관한 연구)

  • Bahn, Gab-su;Mo, Chang-ki;Suh, Eui-kwon
    • Journal of the Korean Society of Industry Convergence
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    • v.10 no.1
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    • pp.5-14
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    • 2007
  • It is necessary for development of drawing product with press to have suitable material selection & all process design and the problem during press process has been cleared from judgement of experience & trial and error. Recently we can estimate press process result from computer aided design & FEM. But we can get more reliable result when we can put more precise process variants during FEM. In case of using a drawbead that is used for the material inflow, it is considered for us to put material property, other analysis condition & friction figure when material is passing through the drawbead for better FEM. From our study, we have drawn an analogy bead connection depth, friction figure & drawing and restraining load according to kinds of lubrication from experiment & FEM for the drawbead. We applied above result to the drawing experiment & FEM and confirmed the validity. We could notice the relation between friction figure & drawing load and the friction figure variation according to kinds of lubrication. It is expected to draw more precise analogy that can be used for real process due to more precise process variants application to FEM.

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Phase Transition Characteristics in $Ge_xSb_{100-x}$ Film for Optical Storage Media

  • Park Tae-jin;Kang Myung-jin;Choi Se-young
    • 정보저장시스템학회:학술대회논문집
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    • 2005.10a
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    • pp.124-127
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    • 2005
  • Rewritable optical memory devices such as an CD-RW and DVD+RW are data storage media, which take advantage of the different optical properties in the amorphous and crystalline states of phase change materials. The switching property, structural transformation, transformation kinetics and chemical bindings of $Ge_xSb_{100-x}$($6{\le}x{\le}$34) were studied to investigate the feasibility of applying $Ge_xSb_{100-x}$ alloys in optical memory. The $Ge_xSb_{100-x}$ thin film was deposited by RF magnetron co-sputtering system and phase change characteristics were investigated by X-ray diffraction (XRD), static tester, inductively coupled plasma atomic emission spectrometer (ICP-AES) and atomic force microscopy (AEM). Optimum fiim composition of $Ge_xSb_{100-x}$ was studied and its minimum time fur laser induced crystallization and optical contrast fur phase transition was performed. These results might be correlated with the binding energies between Ge and Sb, and indicate that $Ge_xSb_{100-x}$ have an potential far optical memory applications.

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Ab initio Studies on Acene Tetramers: Herringbone Structure

  • Park, Young-Hee;Yang, Ki-Yull;Kim, Yun-Hi;Kwon, Soon-Ki
    • Bulletin of the Korean Chemical Society
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    • v.28 no.8
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    • pp.1358-1362
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    • 2007
  • The structures, energetics and transfer integrals of the acene tetramers up to pentacene are investigated with the ab initio molecular orbital method at the level of second-order Møller-Plesset perturbation theory (MP2). Calculated geometries for the herringbone-style structures found in the crystal structure were characterized as local minima, however the geometrical discrepancy between crystal and MP2 theoretical structure is reasonably small. The binding energy of pentacene tetramer was calculated up to 40 kcal/mol (MP2/6-31G(d)) and about 90 kcal/mol (MP2/aug-cc-pVDZ), and the latter seems to be too much overestimated. The tendency of the hole transfer integrals computed with ab initio MP2/3-21G(d) geometry is well agreement with those estimated with crystal structure with some discrepancy, and the gradual increment of the transfer integrals at the crystal geometry is attributed to mainly packing structure rather than the intrinsic property of acene such as a size of acene.

Physicochemical Properties of Sodium Hypochlorite Oxidized Potato Starch (Sodium Hypochlorite로 산화된 감자 전분의 이화학적 특성)

  • 김미라
    • Journal of the East Asian Society of Dietary Life
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    • v.7 no.3
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    • pp.315-324
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    • 1997
  • Physicochemical properties of potato starches oxidized with sodium hypochlorite containing 1.5, 3.0, 4.5, and 6.0% active Cl/g starch at pH 7.0 were examined. Carboxyl group contents of oxidized potato starches were proportional to active chlorine concentration in use and water binding capacity, alkali number, and solubility increased with increasing the degree of oxidation. Blue value and iodine binding property indicated the change of amylose structure by the oxidation. Gelatinization trends obtained from transmittance and DSC thermograms showed that gelatinization temperature was lower as starch was oxidized higher. X-ray diffraction patterns and scanning electron micrographs implied oxidation might occur on the surface amorphous regions of starch granule.

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Computational Study on the Conformational Characteristics of Calix[4]pyrrole Derivatives

  • Hong, Joo-Yeon;Son, Min-Kyung;Ham, Si-Hyun
    • Bulletin of the Korean Chemical Society
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    • v.30 no.2
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    • pp.423-428
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    • 2009
  • The comparative study of three calix[4]heterocycles (calix[4]pyrrole, calix[4]furan, and calix[4]thiophene) has been theoretically performed by using high-level density functional theory (DFT) at the MPWB1K/6-311G$^{**}$//B3LYP/6- 311G$^{**}$ level. The effect of different hetero-atoms (nitrogen, oxygen, and sulfur) placed in the heterocycles on the conformational flexibility, thermodynamic stability order, cavity sizes, charge distributions, and binding propensities are examined. The thermodynamic stability differences between the conformers are found to be much greater in calix[4]pyrrole compared to those in calix[4]furan and calix[4]thiophene. Relatively larger NH group and higher dipole of a pyrrole ring in calix[4]pyrrole contribute to the higher energy barrier for the conformational conversions and relatively rigid potential energy surface compared to the case of calix[4]furan and calix[4]thiophene. The computational results herein provide theoretical understanding of the conformational flexibility and the thermodynamic nature which can be applied to understand the complexation behavior of the three calix[4]heterocycles.

Regulatory Mechanism of L-Alanine Dehydrogenase from Bacillus subtilis

  • Kim, Su Ja;Kim, Yu Jin;Seo, Mi Ran;Jeon, Bong Suk
    • Bulletin of the Korean Chemical Society
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    • v.21 no.12
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    • pp.1217-1221
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    • 2000
  • L-alanine dehydrogenase from Bacillus subtilis exhibits allosteric kinetic properties in the presence of $ZN^{2+}$. $ZN^{2+}$ induces the binding of substrate (L-alanine) to be cooperative at pH 8.0. The effect of pH variation between pH 7.0 and pH 10.0 on the inhibition by $ZN^{2+}$ correlates with the pH effect on the $K_m$ values for L-alanine within these pH range indicating that $ZN^{2+}$ and substrate compete for the same site. No such cooperativity is induced by $ZN^{2+}$ when the reaction is carried out at pH 10. At this higher pH, $ZN^{2+}$ binds with the enzyme with lower affinity and noncompetitive with respect to L-alanine. Inhibition of L-alanine dehydrogenase by $ZN^{2+}$ depends on the ionic strength. Increase in KCI concentration reduced the inhibition, but allosteric property in $ZN^{2+}$ binding is conserved. A model for the regulatory mechanism of L-alanine dehydrogenase as a noncooperative substrate-cooperative cofactor allosteric enzyme, which is compatible in both concerted and the sequential allosteric mechanism, is proposed.

Preparation of $\beta$-Cyclodextrinized Cellulosic Fiber and Deodouring Property ($\beta$-시클로덱스트린화 셀룰로오스 섬유의 제조 및 소취성)

  • Choi, Chang-Nam;Hwang, Tae-Yeon;Ko, Bong-Kook;Kim, Ryong;Hong, Sung-Hak;Kim, Sang-Yool
    • Polymer(Korea)
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    • v.25 no.5
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    • pp.635-641
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    • 2001
  • $\beta$-Cyclodextrine/benzoic acid complex was prepared and reacted with cyanuric chloride (2,4,6-trichloro-1,3,5-triazine). Identification of complex formation and reaction was checked by FT-IR, UV-Vis, and EDX. By reacting this material with cotton fiber, the deodourant fiber was prepared. The deodourizing property was evaluated by the concentration changes of aqueous ammonia solution after flowing ammonia gas through the column titled with deodourant fiber prepared. The deodourizing property was increased with an increase of concentration of $\beta$-cyclodextrine unit in the fiber. In the case of $\beta$-cyclodextrine/benzoic acid complex, the deodourzing property was much increased, comparing with the $\beta$-cyclodextrine only. It was considered to be the binding of aamonia gas caused by benzoic acid in the complex.

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New Unsymmetric Dinuclear Copper(II) Complexes of Trans-disubstituted Cyclam Derivatives: Spectral, Electrochemical, Magnetic, Catalytic, Antimicrobial, DNA Binding and Cleavage Studies

  • Prabu, R.;Vijayaraj, A.;Suresh, R.;Jagadish, L.;Kaviyarasan, V.;Narayanan, V.
    • Bulletin of the Korean Chemical Society
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    • v.32 no.5
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    • pp.1669-1678
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    • 2011
  • Six new binuclear copper(II) complexes have been prepared by template condensation of the dialdehydes 1,8-[bis(3-formyl-2-hydroxy-5-methyl)benzyl]-l,4,8,11-tetraazacyclotetradecane (PC-a) and 1,8-[bis(3-formyl-2-hydroxy-5-bromo)benzyl]-l,4,8,11-tetraazacyclotetradecane (PC-b) with appropriate aliphatic diamines, and copper(II) perchlorate. The structural features of the complexes have been confirmed by elemental analysis, IR, UV-vis and mass spectra etc. The electrochemical behavior of all the copper(II) complexes show two irreversible one electron reduction process. The room temperature magnetic moment studies depict the presence of an antiferromagnetic interaction in the binuclear complexes. The catechol oxidation and hydrolysis of 4-nitrophenylphosphate were carried out by using the complexes as catalyst. The antimicrobial screening data show good results. The binding of the complexes to calf thymus DNA (CT DNA) has been investigated with absorption and emission spectroscopy. The complex [$Cu_2L^{1a}$] displays significant cleavage property of circular plasmid pBR322 DNA in to linear form. Spectral, electrochemical, magnetic and catalytic studies support the distortion of the copper ion geometry that arises as the macrocyclic ring size increases.