• Title/Summary/Keyword: circular dichroism

Search Result 256, Processing Time 0.022 seconds

Soft x-ray Synchrotron Radiation Spectroscopy Study of Molecule-based Nano Bioparticles Containing Fe (철원소를 함유한 분자기반 생체물질 나노입자들의 연 x선 방사광 분광 연구)

  • Lee, Eun-Sook;Kim, D.H.;Hwang, Ji-Hoon;Lee, Ki-Ho;Kang, J.S.
    • Journal of the Korean Magnetics Society
    • /
    • v.22 no.4
    • /
    • pp.125-129
    • /
    • 2012
  • By employing soft x-ray absorption spectroscopy (XAS) and soft x-ray magnetic circular dichroism (XMCD), the electronic structures of molecule-based nano bioparticles, such as Helicobacter pylori ferritin (H. pylori ferritin), Heme, $NaM[Fe(CN)_6]{\cdot}H_2O$-type Prussian Blue (M=Co, Ni) analogue, have been investigated. The measured Fe 2p XAS spectra reveal that Fe ions are trivalent ($Fe^{3+}$) in H. pylori ferritins, while they are in the $Fe^{2+}-Fe^{3+}$ mixed-valent states in $NaM[Fe(CN)_6]{\cdot}H_2O$ Prussian Blue analogues (M=Co, Ni). According to the Fe 2p XMCD spectrum of high-state H. pylori ferritin, all the $Fe^{3+}$ ions have the same local symmetry and their magnetic moments are aligned in the same direction. It is also found that the Fe 3d orbitals in $NaM[Fe(CN)_6]{\cdot}H_2O$ have a strong covalent bonding to $(CN)^-$ ligands, but with a very weak bonding to the 2p orbitals of O ligands.

Differences in the Electronic Structures of Bulk and Powder FeV2O4 Spinel Oxide Investigated by Using Synchrotron Radiation (방사광을 이용한 FeV2O4 스피넬 산화물의 덩치상태와 분말상태의 전자구조 차이 연구)

  • Hwang, Ji-Hoon;Kim, D.H.;Lee, Eun-Sook;Kang, J.S.;Kim, W.C.;Kim, C.S.;Han, S.W.;Hong, S.C.;Park, B.G.;Kim, J.Y.
    • Journal of the Korean Magnetics Society
    • /
    • v.21 no.6
    • /
    • pp.198-203
    • /
    • 2011
  • The electronic structure of ferrimagnetic spinel oxide of $FeV_2O_4$ has been investigated by employing soft x-ray absorption spectroscopy (XAS) and soft x-ray magnetic circular dichroism (XMCD). The Fe 2p and V 2p XAS spectra show that the valence states of Fe and V ions are ${\sim}Fe^{2.3+}$ mixed-valent states and ${\sim}V^{3+}$ states, respectively. In Fe 2p XMCD spectra, finite XMCD signals are observed for divalent $Fe^{2+}$ states only, but not for $Fe^{3+}$ states. This finding indicates that the magnetic moments of $Fe^{2+}$ ions are ordered ferromagnetically but that those of $Fe^{3+}$ ions are cancelled, implying that $Fe^{2+}$ ions play an important role in determining magnetic properties of $FeV_2O_4$.

Complexation of Amphotericin B With Egg Phosphatidylcholine Liposomes

  • Kim, Jin-Chul;Lee, Eun-Ok;Yang, Ji-Won;Choe, Tae-Boo;Kim, Jong-Duk
    • Archives of Pharmacal Research
    • /
    • v.18 no.2
    • /
    • pp.84-89
    • /
    • 1995
  • The complexation and physical characteristics of egg phosphatidylcholine (PC) liposome containing amphotericin B(AmB) were investigated through circular dichrosim(CD) spectra, the size distribution, the turbidity change, and the calcein release. CD spectra of AmB-containing egg PC mxture exhibited a positive peak around 330 nm indicative of complexation of AmB and four negative peaks. The positive peak increased up to $2.2{\;}millidegree/{\mu}g$ AmB as AmB contents increased up to 12% (w/w), suggesting that AmB-phospholipid complexation was promoted by the antibiotics. The effective diameter of liposomesa by dynamic light scattering decreased from 450 nm to 220 nm as the amount of AmB in liposomes increased from o to 30% (w/w). The complexation may be responsible for the reduction in size. On the other hand, at around 1 mN deoxycholate (DOC), the reltive turbidities of 5 and 10% (w/w) AmB-containing liposome suspension were less than 1 probably due to the soblubilization of the complex, while those of pure PC liposome suspension were larger than 1 at the same concentration. Deoxycholate-induced release of liposomes, indicating the intercalation of the drug into the bilayers. Therefore, it is concluded that in AmB/eggPC/water system, AmB-phospholipid complexcoexists with AmB-containing liposomes.

  • PDF

Interaction between Poly(L-lysine) and Poly(N-isopropyl acrylamide-co-acrylic acid) in Aqueous Solution

  • Sung, Yong-Kiel;Yoo, Mi-Kyong;Cho, Chong-Su
    • Macromolecular Research
    • /
    • v.8 no.1
    • /
    • pp.26-33
    • /
    • 2000
  • A series of pH/temperature sensitive polymers were synthesized by copolymerizing N-isopro-pyl acrylamide(NIPAAm) and acrylic acid(AAc) . The influence of polyelectrolyte between poly(allyl amine) (PAA) and poly(L-lysine)(PLL) on the lower critical solution temperature(LCST) of pH/temperature sensitive polymer was compared in the range of pH 2∼12. The LCST of PNIPAAm/water in aqueous poly(NIPAAm-co-AAc) solution was determined by cloud point measurements. A polyelectrolyte complex was prepared by mixing poly(NIPAAm-co-AAc) with poly(allyl amine) (PAA) or poly(L-lysine) (PLL) solutions as anionic and cationic polyelectrolytes, respectively. The effect of polyelectrolyte complex formation on the conformation of PLL was studied as a function of temperature by means of circular dichroism(CD). The cloud points of PNIPAAm in the aqueous copolymers solutions were stongly affected by pH, the presence of polyelectrolyte solute, AAc content, and charge density. The polyelectrolyte complex was formed at neutral condition. The influence of more hydrophobic PLL as a polyelectrolyte on the cloud point of PNIPAAm in the aqueous copolymer solution was stronger than that of poly(allyl amine)(PAA). Although polymer-polymer complex was formed between poly(NIPAAm-co-AAc) and PLL, the conformational change of PLL did not occur due to steric hinderance of bulky N-isopropyl groups of PNIPAAm.

  • PDF

Effect of Carotenoides on the in vitro Aggregation of Bacteriochlorophyll e

  • Hirabayashi, Hiroki;Ohmura, Satoshi;Ishii, Takasada;Uehara, Kaku
    • Journal of Photoscience
    • /
    • v.9 no.2
    • /
    • pp.370-372
    • /
    • 2002
  • In order to investigate effect of the carotenoids (Car) on aggregation of Bacterochlorophyll (BChl) in chlorosome, we studied the spectral difference in aggregates of BChl e formed in the absence and presence of a few kinds of Car in dimethyl sulfoxide (DMSO) -water solution. The absorption spectra of aggregates made of only BChl e and those made of a mixture of BChl e and Car were almost the same. However, the kinetics and circular dichroism (CD) spectra of aggregate of these were markedly different by kind of Car. Specifically, the rate of aggregation for a mixture of BChl e and isorenietene that contains phenyl as end groupe was faster than that for only BChl e. CD spectra of aggregates made of a mixture of BChl e and isorenietene dramatically changed compared to that made of only BChl e. We propose that BChl might form several kinds of rod-like supramolecular structures to in the presence of some kind of Car in chlorosome.

  • PDF

Construction of Two Metal-ion Binding Sites to Improve the 3′-5′Exonuclease Activity of Taq DNA Polymerase

  • Park, Yong-Hyun;Kim, Jong-Moon;Choi, Hye-Ja;Kim, Seog-K.;Kim, Young-Soo
    • Journal of Microbiology and Biotechnology
    • /
    • v.8 no.5
    • /
    • pp.471-477
    • /
    • 1998
  • Taq DNA polymerase from Thermus aquaticus is very useful in the polymerase chain reaction. Taq DNA polymerase is classified in the pol I family, represented by E. coli DNA polymerase I. The three-dimensional structural alignment of 3'-5'exonuclease domains from the pol I family DNA polymerases explains why Taq DNA polymerase does not carry out proofreading in polymerase chain reactions. Three sequence motifs, Exo I, II, and III, must exist to carry out 3'-5'exonuclease activity for proof- reading by a 3'-5'exonuclease reaction, but these are abolished in Taq DNA polymerase. The key catalytic module in 3'-5'exonuclease is two metal ions chelated by four active-site carboxylic amino acids. Taq DNA polymerase was mutagenized to construct the catalytic module in the active site. The circular dichroism technique supported the formation of the catalytic module, and the radioactive assay showed that the 3'-5'exonuclease activity doubled in the mutant Taq DNA polymerase.

  • PDF

Non-Essential Activation of Co2+ and Zn2+ on Mushroom Tyrosinase: Kinetic and Structural Stability

  • Gheibi, N.;Saboury, A.A.;Sarreshtehdari, M.
    • Bulletin of the Korean Chemical Society
    • /
    • v.32 no.5
    • /
    • pp.1500-1506
    • /
    • 2011
  • Tyrosinase is a widespread enzyme with great promising capabilities. The Lineweaver-Burk plots of the catecholase reactions showed that the kinetics of mushroom tyrosinase (MT), activated by $Co^{2+}$ and $Zn^{2+}$ at different pHs (6, 7, 8 and 9) obeyed the non-essential activation mode. The binding of metal ions to the enzyme increases the maximum velocity of the enzyme due to an increase in the enzyme catalytic constant ($k_{cat}$). From the kinetic analysis, dissociation constants of the activator from the enzyme-metal ion complex ($K_a$) were obtained as $5{\times}10^4M^{-1}$ and $8.33{\times}10^3M^{-1}$ for $Co^{2+}$ and $Zn^{2+}$ at pH 9 and 6 respectively. The structural analysis of MT through circular dichroism (CD) and intensive fluorescence spectra revealed that the conformational stability of the enzyme in these pHs reaches its maximum value in the presence of each of the two metal ions.

Biological Characterization of the Omp1-like Protein from Actinobacillus actinomycetemcomitans

  • Ha, Jung-Hye;Jeong, Mi-Suk;Jo, Wol-Soon;Jeong, Min-Ho;Jang, Se-Bok
    • Bulletin of the Korean Chemical Society
    • /
    • v.31 no.2
    • /
    • pp.275-280
    • /
    • 2010
  • Actinobacillus actinomycetemcomitans is a gram-negative, nonmotile coccobacillus bacterium that is associated with several human diseases, including endocarditis, meningitis, osteomyelitis, subcutaneous abscesses and periodontal diseases. A full-length Omp1-like protein gene from A. actinomycetemcomitans was cloned into a pQE30 vector and overexpressed in Escherichia coli BL21(DE3) cells. The protein revealed sequence homologies to Seventeen kilodalton proteins (Skp) from Pasteurella multocida and E. coli that have been characterized as periplasmic chaperones. This soluble Omp1-like protein was successfully purified to homogeneity for further folding and functional studies. The purity, identity, and conformation of the protein were determined using sodium dodecyl sulfate polyacrylamide gel electrophoresis, matrix-assisted laser desorption ionization mass spectrometry, circular dichroism, fluorescence spectroscopic, and differential scanning calorimetric studies. We showed that the protein formed an oligomer larger than a tetramer. We found, further, that it is comprised of mostly $\alpha$-helices and boasts high thermal stability.

Conformational and Structural Changes of Choline Oxidase from Alcaligenes Species by Changing pH Values

  • Hekmat, A.;Saboury, A. A.;Divsalar, A.;Khanmohammadi, M.
    • Bulletin of the Korean Chemical Society
    • /
    • v.29 no.8
    • /
    • pp.1510-1518
    • /
    • 2008
  • Results of intrinsic and extrinsic fluorescence studies on choline oxidase revealed that the enzyme at high alkaline pH values has more accessible hydrophobic patches relative to acidic pH. Fluorescence quenching studies with acrylamide confirm these changes. The quenching constants were also determined at different pH(s) by using the Stern-Volmer equation. CD studies showed that at higher pH a transition from $\alpha$-helix to $\beta$- structure was appeared while at lower pH the content of $\alpha$-helix structure was increased. Furthermore, analysis of the spectral data using chemometric method gave evidence for existence of intermediate components at very high pH(s). Results of thermal denaturation evaluated that the enzyme has the most instability at higher pH(s). Altogether low and high pH values caused significant alteration on secondary and tertiary structures of choline oxidase via inducing of an intermediate.

PLGA-Loaded Gold-Nanoparticles Precipitated with Quercetin Downregulate HDAC-Akt Activities Controlling Proliferation and Activate p53-ROS Crosstalk to Induce Apoptosis in Hepatocarcinoma Cells

  • Bishayee, Kausik;Khuda-Bukhsh, Anisur Rahman;Huh, Sung-Oh
    • Molecules and Cells
    • /
    • v.38 no.6
    • /
    • pp.518-527
    • /
    • 2015
  • Controlled release of medications remains the most convenient way to deliver drugs. In this study, we precipitated gold nanoparticles with quercetin. We loaded gold-quercetin into poly(DL-lactide-co-glycolide) nanoparticles (NQ) and tested the biological activity of NQ on HepG2 hepatocarcinoma cells to acquire the sustained release property. We determined by circular dichroism spectroscopy that NQ effectively caused conformational changes in DNA and modulated different proteins related to epigenetic modifications and c ell cycle control. The mitochondrial membrane potential (MMP), reactive oxygen species (ROS), cell cycle, apoptosis, DNA damage, and caspase 3 activity were analyzed by flow cytometry, and the expression profiles of different anti- and pro-apoptotic as well as epigenetic signals were studied by immunoblotting. A cytotoxicity assay indicated that NQ preferentially killed cancer cells, compared to normal cells. NQ interacted with HepG2 cell DNA and reduced histone deacetylases to control cell proliferation and arrest the cell cycle at the sub-G stage. Activities of cell cycle-related proteins, such as $p21^{WAF}$, cdk1, and pAkt, were modulated. NQ induced apoptosis in HepG2 cells by activating p53-ROS crosstalk and induces epigenetic modifications leading to inhibited proliferation and cell cycle arrest.