• 제목/요약/키워드: affinity interaction

검색결과 240건 처리시간 0.027초

Interaction of Resveratrol and Genistein with Nucleic Acids

  • Usha, Subbiah;Johnson, Irudayam Maria;Malathi, Raghunathan
    • BMB Reports
    • /
    • 제38권2호
    • /
    • pp.198-205
    • /
    • 2005
  • Resveratrol (RES) and genistein (GEN) are the dietary natural products known to possess chemopreventive property and also the ability to repair DNA damage induced by mutagens/carcinogens. It is believed that the therapeutic activity of these compounds could be primarily due to their interaction with nucleic acids but detailed reports are not available. We here explore the interaction of these drugs with nucleic acids considering DNA and RNA as a potential therapeutic target. The interaction of RES and GEN has been analysed in buffered solution with DNA [saline sodium citrate (SSC)] and RNA [tris ethylene diammine tetra acetic acid (TE)] using UV-absorption and Fourier transform infrared (FTIR) spectroscopy. The UV analysis revealed lesser binding affinity with nucleic acids at lower concentration of RES (P/D = 5.00 and 10.00), while at higher drug concentration (P/D = 0.75, 1.00 and 2.50) hyperchromic effect with shift in the ${\lambda}_{max}$ is noted for DNA and RNA. A major RES-nucleic acids complexes was observed through base pairs and phosphate backbone groups with K = $35.782\;M^{-1}$ and K = $34.25\;M^{-1}$ for DNA-RES and RNA-RES complexes respectively. At various concentrations of GEN (P/D = 0.25, 0.50, 0.75, 1.00 and 2.50) hyperchromicity with shift in the ${\lambda}_{max}$ from 260 $\rightarrow$ 263 om and 260 $\rightarrow$ 270 nm is observed for DNA-GEN and RNA-GEN complexes respectively. The binding constant (from UV analysis) for GEN-nucleic acids complexes could not be obtained due to GEN absorbance overlap with that of nucleic acids at 260 nm. Nevertheless a detailed analysis with regard to the interaction of these drugs (RES/GEN) with DNA and RNA could feasibly be understood by FTIR spectroscopy. The NH band of free DNA and RNA which appeared at $3550-3100\;cm^{-1}$ and $3650-2700\;cm^{-1}$ shifted to $3450-2950\;cm^{-1}$ and $3550-3000\;cm^{-1}$ in DNA-RES and RNA-RES complexes respectively. Similarly shifts corresponding to $3650-3100\;cm^{-1}$ and $3420-3000\;cm^{-1}$ have been observed in DNA-GEN and RNA-GEN complexes respectively. The observed reduction in NH band of free nucleic acids upon complexation of these drugs is an indication of the involvement of the hydroxyl (OH) and imino (NH) group during the interaction of the drugs and nucleic acids (DNA/RNA) through H-bonded formation. The interaction of RES and GEN with bases appears in the order of G $\geq$ T > C > A and A > C $\geq$ T > G. Further interaction of these natural compounds with DNA and RNA is also supported by changes in the vibrational frequency (shift/intensity) in symmetrical and asymmetrical stretching of aromatic rings of drugs in the complex spectra. No appreciable shift is observed in the DNA and RNA marker bands, indicating that the B-DNA form and A-family conformation of RNA are not altered during their interaction with RES and GEN.

Drug-Biomacromolecule Interaction VIII

  • Kim, Chong-Kook;Yang, Ji-Sun;Lim, Yun-Su
    • Archives of Pharmacal Research
    • /
    • 제7권2호
    • /
    • pp.87-93
    • /
    • 1984
  • The effects of ionic strength and pH on the binding of cefazolin to bovine serum albumin (BSA) were studied by UV difference spectrophotometry. As ionic strength at constant pH and temperature increases, the apparent bining constant decreased but the number of binding sites remained almost constant at 2. The constancy of the number of binding sites with increasing the ionic strength suggests that purely electrostatic forces between BSA and drug do not have great importance in the drug binding, even though there is a decrease in the apparent binding constant. Thus, the effect of ionic strength on the interaction between drug and BSA may be explained by the changes in ionic atmosphere of the aggregated BSA molecules and competitive inhibition by phosphate ions. In addition, the higher apparent binding constant at high ionic strength is explained by conformational changes of BSA from its aggregate forms into subunits. The pH effects on the afinity of interactions indicated that the binding affinity of cefazoline is higher in the neutral region than in the alkaline region. An d at high pH value, the number of binding sites decreased from 2 to 1 because of the conformational change of BSA in the alkaline region.

  • PDF

In vitro Evidence that Purified Yeast Rad27 and Dna2 are not Stably Associated with Each Other Suggests that an Additional Protein(s) is Required for a Complex Formation

  • Bae, Sung-Ho;Seo, Yeon-Soo
    • BMB Reports
    • /
    • 제33권2호
    • /
    • pp.155-161
    • /
    • 2000
  • The saccharomyces cerevisiae Rad27, a structure-specific endonuclease for the okazaski fragment maturation has been known to interact genetically and biochemically with Dna2, an essential enzyme for DNA replication. In an attempt to define the significance of the interaction between the two enzymes, we expressed and purified both Dna2 and Rad27 proteins. In this report, Rad27 could not form a complex with Dna2 in the three different analyses. The analyses included glycerol gradient sedimentation, protein-column chromatography, and coinfection of baculoviruses followed by affinity purification. This is in striking contrast to the previous results that used crude extracts. These results suggest that the interaction between the two proteins is not sufficiently stable or indirect, and thus requires an additional protein(s) in order for Rad27 and Dna2 to form a stable physical complex. This result is consistent with our genetic findings that Schizosaccharomyces pombe Dna2 is capable of interacting with several proteins that include two subunits of polymerase $\delta$, DNA ligase I, as well as Fen-1. In addition, we found that the N-terminal modification of Rad27 abolished its enzymatic activity. Thus, as suspected, we found that on the basis of the structure determination, N-terminal methionine indeed plays an important role in the nucleolytic cleavage reaction.

  • PDF

Functional Regulation of Dopamine D3 Receptor through Interaction with PICK1

  • Zheng, Mei;Zhang, Xiaohan;Min, Chengchun;Choi, Bo-Gil;Oh, In-Joon;Kim, Kyeong-Man
    • Biomolecules & Therapeutics
    • /
    • 제24권5호
    • /
    • pp.475-481
    • /
    • 2016
  • PICK1, a PDZ domain-containing protein, is known to increase the reuptake activities of dopamine transporters by increasing their expressions on the cell surface. Here, we report a direct and functional interaction between PICK1 and dopamine $D_3$ receptors ($D_3R$), which act as autoreceptors to negatively regulate dopaminergic neurons. PICK1 colocalized with both dopamine $D_2$ receptor ($D_2R$) and $D_3R$ in clusters but exerted different functional influences on them. The cell surface expression, agonist affinity, endocytosis, and signaling of $D_2R$ were unaffected by the coexpression of PICK1. On the other hand, the surface expression and tolerance of $D_3R$ were inhibited by the coexpression of PICK1. These findings show that PICK1 exerts multiple effects on $D_3R$ functions.

The Role of Dendritic Cells in Central Tolerance

  • Oh, Jaehak;Shin, Jeoung-Sook
    • IMMUNE NETWORK
    • /
    • 제15권3호
    • /
    • pp.111-120
    • /
    • 2015
  • Dendritic cells (DCs) play a significant role in establishing self-tolerance through their ability to present self-antigens to developing T cells in the thymus. DCs are predominantly localized in the medullary region of thymus and present a broad range of self-antigens, which include tissue-restricted antigens expressed and transferred from medullary thymic epithelial cells, circulating antigens directly captured by thymic DCs through coticomedullary junction blood vessels, and peripheral tissue antigens captured and transported by peripheral tissue DCs homing to the thymus. When antigen-presenting DCs make a high affinity interaction with antigen-specific thymocytes, this interaction drives the interacting thymocytes to death, a process often referred to as negative selection, which fundamentally blocks the self-reactive thymocytes from differentiating into mature T cells. Alternatively, the interacting thymocytes differentiate into the regulatory T (Treg) cells, a distinct T cell subset with potent immune suppressive activities. The specific mechanisms by which thymic DCs differentiate Treg cells have been proposed by several laboratories. Here, we review the literatures that elucidate the contribution of thymic DCs to negative selection and Treg cell differentiation, and discusses its potential mechanisms and future directions.

Modulation of the Cytochrome c Oxidase Activity by ATP: Implications for Mitochondrial Respiratory Control

  • Park, Nan-Hyang;Chun, Sun-Bum;Han, Tae-Young;Han, Sang-Hwa
    • BMB Reports
    • /
    • 제29권4호
    • /
    • pp.300-307
    • /
    • 1996
  • ATP and ADP are potential regulators of mitochondtial respiration and at physiological concentrations they affect the rate of electron transfer between cytochrome c and cytochrome c oxidase. The electron transfer, however, depends on the electrostatic interaction between the two proteins. In order to exclude any nonspecific ionic effects by these polyvalent nucleotides, we used 2'-O-(2,4,6)trinitro(TNP)-derivatives of ATP and ADP which have three orders of magnitude higher affinity for cytochrome c oxidase. A simple titration of the fluorescence intensity of TNP by cytochrome c oxidase showed a binding stoichiometry of 2:1 cytochrome c:cytochrome c oxidase. Higher ionic strength was required for TNP-ATP than for TNP-ADP to be dissociated from cytochrome c oxidase, indicating that the negative charges on the phosphate group are at least partially responsible for the binding. In both spectrophotometric and polarographic assays, addition of ATP (and ADP to a less extent) showed an enhanced cytochrome c oxidase activity. Both electron paramagnetic resonance and fluorescence spectra indicate that there is no Significant change in the cytochrome c-cytochrome c oxidase interaction. Instead, reduction levels of the cytochromes at steadystate suggest that the increased activity of nucleotide-bound cytochrome c oxidase is due to faster electron transfer from cytochrome ${\alpha}$ to cytochrome ${\alpha}_3$, which is known to be the fate limiting step in the oxygen reduction by cytochrome c oxidase.

  • PDF

고감도 나노-바이오센서를 위한 나노로드 전극 표면 개질에 관한 연구 (A Study on Surface Modification of Nanorod Electrodes for Highly Sensitive Nano-biosensor)

  • 이승준
    • 공업화학
    • /
    • 제27권2호
    • /
    • pp.185-189
    • /
    • 2016
  • 많은 생체 친화적인 센서들 중에서 avidin-biotin system은 높은 상호 특이적인 친화성으로 인하여 많은 생물학적인 응용 연구에 이용되어 왔다. 효과적인 avidin-biotin 바이오센서 개발을 위해 avidin-biotin 간의 상호 반응성을 증대시키기 위해서는 높은 표면적을 가지는 전극이 필요하다. 본 연구에서는 이러한 목적을 위해 gold nanorods electrode를 사용하였다. 전기화학적인 특성은 cyclic voltammetry (CV)와 electrochemical impedance spectroscopy (EIS)를 가지고 redox couple $[Fe(CN)_6]^{3-/4-}$를 사용하여 다양한 biotin의 농도에 따라 분석되었다. 결론적으로 nanorod의 전극은 1 ng/mL보다 낮은 biotin의 농도도 감지할 수 있음을 보였다.

Removal of Methylene blue from saline solutions by adsorption and electrodialysis

  • Lafi, Ridha;Mabrouk, Walid;Hafiane, Amor
    • Membrane and Water Treatment
    • /
    • 제10권2호
    • /
    • pp.139-148
    • /
    • 2019
  • In this study, the removal of MB from saline solutions was evaluated by two methods by adsorption and electrodialysis; the adsorption of the mixture dye/salt on dried orange peel waste (OPW) was studied in batch method. In this study the biosorption of cationic dye by OPW was investigated as a function of initial solution pH, and initial salt (sodium chloride) concentration. The maximal dye uptake at $pH{\geq}3.6$ in the absence and in the presence of salt and the dye uptake diminished considerably in the presence of increasing concentrations of salt up to 8 g/L. The Redlich Peterson and Langmuir were the most suitable adsorption models for describing the biosorption equilibrium data of the dye both individually and in salt containing medium. As well, this work deals with the electrodialysis application to remove the dye. Synthetic solutions were used for the investigation of the main operational factors affecting the treatment performance; such as applied voltage, pH, initial dye concentration and ionic strength. The experimental results for adsorption and electrodialysis confirmed the importance of electrostatic interactions on the dye. The electrodialysis process with standard ion exchange membranes enabled efficient desalination of cationic dye solutions; there are two main factors in fouling: electrostatic interaction between cations of dyes and the fixed charged groups of the CEM, and affinity interactions.

Coil-to-globule transition of thermo-responsive γ-substituted poly (ɛ-caprolactone) in water: A molecular dynamics simulation study

  • Koochaki, Amin;Moghbeli, Mohammad Reza;Nikkhah, Sousa Javan
    • Current Applied Physics
    • /
    • 제18권11호
    • /
    • pp.1313-1319
    • /
    • 2018
  • The coil-to-globule behavior of poly{${\gamma}$-2-[2-(2methoxyethoxy)ethoxy]ethoxy-3-caprolactone} (PMEEECL) as a ${\gamma}$-substituted poly (${\varepsilon}$-caprolactone) was investigated via atomistic molecular dynamics (MD) simulation. For this purpose, radius of gyration, end-to-end distance and radial distribution function of the chain in the presence of water were calculated. Consequently, the lower critical solution temperature (LCST) of PMEEECL chain at which the coil-to-globule transition takes place, was determined in each calculated parameter curve. The simulation results indicated that the LCST of PMEEECL was occurred at close to 320 K, which is in a good agreement with previous experimental results. Additionally, the appearance of sudden change in both Flory-Huggins interaction parameter (${\chi}$) and interaction energy between the PMEEECL chain and water molecules at about 320 K confirmed the calculated LCST result. The radial distribution function (RDF) results showed that the affinity of the PMEEECL side chain to water molecules is lower than its backbone.

Molecular Dynamics Simulation of Adhesion Processes

  • Cho, Sung-San;Park, Seungho
    • Journal of Mechanical Science and Technology
    • /
    • 제16권11호
    • /
    • pp.1440-1447
    • /
    • 2002
  • Adhesion of a hemispherical tip to the flat surface in nano-structures is simulated using the molecular dynamics technique. The tip and plates are modeled with the Lennard-Jones molecules. The simulation focuses on the deformation of the tip. Detailed descriptions on the evolution of interaction force, the energy dissipation due to adhesion hysteresis, the forma- tion-growth-breakage of adhesive junction as well as the evolution of molecular distribution during the process are presented. The effects of the tip size, the maximum tip approach, the tip temperature, and the affinity between the tip and the mating plate are also discussed.