• 제목/요약/키워드: Negatively charged

검색결과 246건 처리시간 0.023초

Comparison of the Binding Modes of [Ru(2,2'-bipyridine)3]2+ and [Ru(2,2':6',2"-terpyridine)2]2+ to Native DNA

  • Jang, Yoon-Jung;Lee, Hyun-Mee;Jang, Kyeung-Joo;Lee, Jae-Cheol;Kim, Seog-K.;Cho, Tae-Sub
    • Bulletin of the Korean Chemical Society
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    • 제31권5호
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    • pp.1314-1318
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    • 2010
  • The $[Ru(tpy)_2]Cl_2$ (tpy:2,2':6',2"-terpyridine) complex was synthesized and its structure was confirmed by $^1H$-NMR and elemental analysis. Its binding mode toward DNA was compared with the well-known $[Ru(bpy)_3]Cl_2$ (bpy:2,2-bipyridyl), using isotropic absorption, linear dichroism(LD) spectroscopy, and an energy minimization study. Compared to $[Ru(bpy)_3]^{2+}$, the $[Ru(tpy)_2]^{2+}$ complex exhibited very little change in its absorption pattern, especially in the MLCT band, upon binding to DNA. Furthermore, upon DNA binding, both Ru(II) complexes induced a decrease in the LD magnitude in the DNA absorption region. The $[Ru(tpy)_2]^{2+}$ complex produced a strong positive LD signal in the ligand absorption region, which is in contrast with the $[Ru(bpy)_3]^{2+}$ complex. Observed spectral properties led to the conclusion that the interaction between the ligands and DNA bases is negligible for the $[Ru(tpy)_2]^{2+}$ complex, although it formed an adduct with DNA. This conclusion implies that both complexes bind to the surface of DNA, most likely to negatively charged phosphate groups via a simple electrostatic interaction, thereby orienting to exhibit the LD signal. The energy minimization calculation also supported this conclusion.

Heat or radiofrequency plasma glow discharge treatment of a titanium alloy stimulates osteoblast gene expression in the MC3T3 osteoprogenitor cell line

  • Rapuano, Bruce E.;Hackshaw, Kyle;Macdonald, Daniel E.
    • Journal of Periodontal and Implant Science
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    • 제42권3호
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    • pp.95-104
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    • 2012
  • Purpose: The purpose of this study was to determine whether increasing the Ti6Al4V surface oxide negative charge through heat ($600^{\circ}C$) or radiofrequency plasma glow discharge (RFGD) pretreatment, with or without a subsequent coating with fibronectin, stimulated osteoblast gene marker expression in the MC3T3 osteoprogenitor cell line. Methods: Quantitative real-time polymerase chain reaction was used to measure changes over time in the mRNA levels for osteoblast gene markers, including alkaline phosphatase, bone sialoprotein, collagen type I (${\alpha}1$), osteocalcin, osteopontin and parathyroid hormone-related peptide (PTH-rP), and the osteoblast precursor genes Runx2 and osterix. Results: Osteoprogenitors began to differentiate earlier on disks that were pretreated with heat or RFGD. The pretreatments increased gene marker expression in the absence of a fibronectin coating. However, pretreatments increased osteoblast gene expression for fibronectin-coated disks more than uncoated disks, suggesting a surface oxide-mediated specific enhancement of fibronectin's bioactivity. Heat pretreatment had greater effects on the mRNA expression of genes for PTH-rP, alkaline phosphatase and osteocalcin while RFGD pretreatment had greater effects on osteopontin and bone sialoprotein gene expression. Conclusions: The results suggest that heat and RFGD pretreatments of the Ti6Al4V surface oxide stimulated osteoblast differentiation through an enhancement of (a) coated fibronectin's bioactivity and (b) the bioactivities of other serum or matrix proteins. The quantitative differences in the effects of the two pretreatments on osteoblast gene marker expression may have arisen from the unique physico-chemical characteristics of each resultant oxide surface. Therefore, engineering the Ti6Al4V surface oxide to become more negatively charged can be used to accelerate osteoblast differentiation through fibronectin-dependent and independent mechanisms.

Structure-activity relationships of cecropin-like peptides and their interactions with phospholipid membrane

  • Lee, Eunjung;Jeong, Ki-Woong;Lee, Juho;Shin, Areum;Kim, Jin-Kyoung;Lee, Juneyoung;Lee, Dong Gun;Kim, Yangmee
    • BMB Reports
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    • 제46권5호
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    • pp.282-287
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    • 2013
  • Cecropin A and papiliocin are novel 37-residue cecropin-like antimicrobial peptides isolated from insect. We have confirmed that papiliocin possess high bacterial cell selectivity and has an ${\alpha}$-helical structure from $Lys^3$ to $Lys^{21}$ and from $Ala^{25}$ to $Val^{35}$, linked by a hinge region. In this study, we demonstrated that both peptides showed high antimicrobial activities against multi-drug resistant Gram negative bacteria as well as fungi. Interactions between these cecropin-like peptides and phospholipid membrane were studied using CD, dye leakage experiments, and NMR experiments, showing that both peptides have strong permeabilizing activities against bacterial cell membranes and fungal membranes as well as $Trp^2$ and $Phe^5$ at the N-terminal helix play an important role in attracting cecropin-like peptides to the negatively charged bacterial cell membrane. Cecropin-like peptides can be potent peptide antibiotics against multi-drug resistant Gram negative bacteria and fungi.

유동흐름 전류계를 이용한 정수장 고탁도 유입수 응집 제어 방법에 대한 연구 (Coagulation Control of High Turbid Water Samples Using a Streaming Current Control System)

  • 남승우;조병일;김원경;조경덕
    • 한국환경보건학회지
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    • 제38권2호
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    • pp.128-135
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    • 2012
  • Objectives: This study was aimed at determining the optimum coagulation dosage in a high turbid kaolin water sample using streaming current detection (SCD) as an alternative to the jar test. Methods: SCD is able to optimize coagulant dosing by titration of negatively charged particles. Kaolin particles were used to mimic highly turbid water ranging from 50 to 600 NTU, and polyaluminum chloride (PAC, 17%) was applied as a titrant and coagulant. The coagulation consisted of rapid stirring (5 min at 140 rpm), reduced stirring (20 min at 70 rpm), and settling (60 min). To confirm the coagulation effect, a jar test was also compared with the SCD titration results. Results: SCD titration of kaolin water samples showed that the dose of PAC increased as the pH rose. However, supernatant turbidity less than 1 NTU after coagulation was not achieved for high turbid water by SCD titration. Instead, a conversion factor was used to calculate the optimum PAC dosage for high turbid water by correlating a jar test result with that from an SCD titration. Using this approach, we were able to successfully achieve less than 1 NTU in treated water. Conclusions: For high turbid water influent in a water treatment plant, particularly during summer, the application of SCD control by applying a conversion factor can be more useful than a jar test due to the rapid calculation of coagulation dosage. Also, the interpolation of converted PAC dose could successfully achieve turbidity in the treated water of less than 1 NTU. This result indicates that an SCD system can be effectively used in a water treatment plant even for high turbid water during the rainy season.

전기화학 바이오센서의 전극물질로 응용을 위한 열분해 탄소의 제작 및 특성 연구 (Fabrication and Characterization of Pyrolyzed Carbon for Use as an Electrode Material in Electrochemical Biosensor)

  • 이정아;황성필;곽주현;박세일;이승섭;이광철
    • 대한기계학회논문집A
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    • 제31권10호
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    • pp.986-992
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    • 2007
  • This paper presents the fabrication and characterization of carbon films pyrolyzed with various photoresists for bioMEMS applications. To verify the usefulness of pyrolyzed carbon films as an electrode material in an electrochemical biosensor developed by the authors, interactions between avidin and biotin on the pyrolyzed carbon film were studied via electrochemical impedance spectroscopy based on electrostatic interactions between avidin and negatively-charged ferricyanide. The pyrolyzed carbon films were characterized using a surface profiler, a precision semiconductor parameter analyzer, a nanoindentor, scanning electron microscopy, and atomic force microscopy. Amine conjugated biotin was immobilized on the electrode using EDC/NHS as crosslinkers after $O_2$ plasma treatment to enhance functional groups on the carbon electrode pyrolyzed at $1000^{\circ}C$ with AZ9260. The detection of avidin binding with different concentrations in a range of 0.75 nM to $7.5\;{\mu}M$ to the pyrolyzed carbon electrode modified with biotin was carried out by measuring the electrochemical impedance change. The results show that avidin binds to the biotin on the electrode not by non-specific interaction but by specific interaction, and that EIS successfully detects this binding event. Pyrolyzed carbon films are a promising material for miniaturization, integration, and low-cost fabrication in electrochemical biosensors.

Potential of Mean Force Calculations for Ion Selectivity in a Cyclic Peptide Nanotube

  • Choi, Kyu-Min;Kwon, Chan-Ho;Kim, Hong-Lae;Hwang, Hyon-Seok
    • Bulletin of the Korean Chemical Society
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    • 제33권3호
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    • pp.911-916
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    • 2012
  • Ion selectivity in a simple cyclic peptide nanotube, composed of four cyclo[-(D-Ala-Glu-D-Ala-Gln)$_2-$] units, is investigated by calculating the PMF profiles of $Na^+$, $K^+$, and $Cl^-$ ions permeating through the peptide nanotube in water. The final PMF profiles of the ions obtained from the umbrella sampling (US) method show an excellent agreement with those from the thermodynamic integration (TI) method. The PMF profiles of $Na^+$ and $K^+$ display free energy wells while the PMF curve of $Cl^-$ features free energy barriers, indicating the selectivity of the cyclic peptide nanotube to cations. Decomposition of the total mean force into the contribution from each component in the system is also accomplished by using the TI method. The mean force decomposition profiles of $Na^+$ and $K^+$ demonstrate that the dehydration free energy barriers by water molecules near the channel entrance and inside the channel are completely compensated for by attractive electrostatic interactions between the cations and carbonyl oxygens in the nanotube. In the case of $Cl^-$, the dehydration free energy barriers are not eliminated by an interaction between the anion and the peptide nanotube, leading to the high free energy barriers in the PMF profile. Calculations of the coordination numbers of the ions with oxygen atoms pertaining to either water molecules or carbonyl groups in the peptide nanotube reveal that the stabilization of the cations in the midplane regions of the nanotube arises from the favorable interaction of the cations with the negatively charged carbonyl oxygens.

Analysis of interaction between intracellular spermine and transient receptor potential canonical 4 channel: multiple candidate sites of negatively charged amino acids for the inward rectification of transient receptor potential canonical 4

  • Kim, Jinsung;Moon, Sang Hui;Kim, Taewook;Ko, Juyeon;Jeon, Young Keul;Shin, Young-Cheul;Jeon, Ju-Hong;So, Insuk
    • The Korean Journal of Physiology and Pharmacology
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    • 제24권1호
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    • pp.101-110
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    • 2020
  • Transient receptor potential canonical 4 (TRPC4) channel is a nonselective calcium-permeable cation channels. In intestinal smooth muscle cells, TRPC4 currents contribute more than 80% to muscarinic cationic current (mIcat). With its inward-rectifying current-voltage relationship and high calcium permeability, TRPC4 channels permit calcium influx once the channel is opened by muscarinic receptor stimulation. Polyamines are known to inhibit nonselective cation channels that mediate the generation of mIcat. Moreover, it is reported that TRPC4 channels are blocked by the intracellular spermine through electrostatic interaction with glutamate residues (E728, E729). Here, we investigated the correlation between the magnitude of channel inactivation by spermine and the magnitude of channel conductance. We also found additional spermine binding sites in TRPC4. We evaluated channel activity with electrophysiological recordings and revalidated structural significance based on Cryo-EM structure, which was resolved recently. We found that there is no correlation between magnitude of inhibitory action of spermine and magnitude of maximum current of the channel. In intracellular region, TRPC4 attracts spermine at channel periphery by reducing access resistance, and acidic residues contribute to blocking action of intracellular spermine; channel periphery, E649; cytosolic space, D629, D649, and E687.

Bioremediation of Diesel-Contaminated Soil by Bacterial Cells Transported by Electrokinetics

  • LEE, HYO-SANG;KISAY LEE
    • Journal of Microbiology and Biotechnology
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    • 제11권6호
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    • pp.1038-1045
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    • 2001
  • The electrokinetic technology was applied in bioremediation for the purpose of supplying a Pseudomonas strain capable of degrading diesel to contaminated soil bed, and their biodegradation of diesel was carried out after a desired cell distribution was obtained. Electrokinetic injection of the strain was made possible because the cells acted as negatively charged particles at neutral pH, and thus the cells were transported with a precise directionality through the soil mostly by the mechanism of electrophoresis and in part by electroosmosis. A severe pH change in the soil bed was formed due to the penetration of electrolysis products, which was harmful to the cell viability and cell transport. To achieve a desirable cell transport and distribution, the control of pH in soil bed by a recirculating buffer solution in electrode chambers was essential during the appliation of an electric field. The judicious selections of electrolyte concentration and conductivity were also important for achieving an efficient electrokinetic cell transport since a higher electrolyte concentration favored the maintenance of pH stability in soil bed, but lowered electrophoretic mobility on the other hand. With electrolyte solution of pH 7 phosphate buffer, a 0.05 M concentration showed a better cell transport buffer, a 0.05 M concentration showed a better cell transport than 0.02 M and 0.08 M. The cell under pH 8 were obtained, compared to the cells under pH 7 or pH 9 in a given time period Up to $60\%$ of diesel was degraded in 8 days by the Pseudomonas cell, which were distributed electrokinetically under the conditions of pH 8 ($1,800{\mu}S/cm$, a mixture of phosphate and ammonia buffers) and 40 mA in a soil bed of 15 cm length.

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Performance of membrane filtration in the removal of iron and manganese from Malaysia's groundwater

  • Kasim, Norherdawati;Mohammad, Abdul Wahab;Abdullah, Siti Rozaimah Sheikh
    • Membrane and Water Treatment
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    • 제7권4호
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    • pp.277-296
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    • 2016
  • The aim of this research was to investigate the ability of nanofiltration (NF) and ultrafiltration (UF) membranes as a filtration unit for groundwater treatment for drinking water resources. Commercial membranes denoted as TS40, TFC-SR3 and GHSP were used to study the performance based on rejections and fluxes. The investigation has been conducted using natural groundwater obtained from a deep tube well with initial concentration of iron (Fe) and manganese (Mn) at 7.15 mg/L and 0.87 mg/L, respectively. Experimental results showed that NF membranes exhibited higher fluxes than UF membrane with pure water permeability at 4.68, 3.99 and $3.15L.m^{-2}.h^{-1}.bar^{-1}$, respectively. For metal rejection, these membranes have performed higher removal on Fe with TS40, TFC-SR3 and GHSP membranes having more than 82%, 92% and 86% respectively. Whereas, removal on Mn only achieved up to 60%, 80% and 30%, for TS40, TFC-SR3 and GHSP membranes respectively. In order to achieve drinking water standard, the membranes were efficient in removing Fe ion at 1 and 2 bar in contrast with Mn ion at 4 and 5 bar. Higher rejection of Fe and Mn were achieved when pH of feed solution was increased to more than 7 as TFC-SR3 membrane was negatively charged in basic solution. This effect could be attributed to the electrostatic effect interaction between membrane material and rejected ions. In conclusion, this study proved that NF membrane especially the TFC-SR3 membrane successfully treated local groundwater sources for public drinking water supply in line with the WHO standard.

Crystal Structure of Cytochrome cL from the Aquatic Methylotrophic Bacterium Methylophaga aminisulfidivorans MPT

  • Ghosh, Suparna;Dhanasingh, Immanuel;Ryu, Jaewon;Kim, Si Wouk;Lee, Sung Haeng
    • Journal of Microbiology and Biotechnology
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    • 제30권8호
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    • pp.1261-1271
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    • 2020
  • Cytochrome cL (CytcL) is an essential protein in the process of methanol oxidation in methylotrophs. It receives an electron from the pyrroloquinoline quinone (PQQ) cofactor of methanol dehydrogenase (MDH) to produce formaldehyde. The direct electron transfer mechanism between CytcL and MDH remains unknown due to the lack of structural information. To help gain a better understanding of the mechanism, we determined the first crystal structure of heme c containing CytcL from the aquatic methylotrophic bacterium Methylophaga aminisulfidivorans MPT at 2.13 Å resolution. The crystal structure of Ma-CytcL revealed its unique features compared to those of the terrestrial homologues. Apart from Fe in heme, three additional metal ion binding sites for Na+, Ca+, and Fe2+ were found, wherein the ions mostly formed coordination bonds with the amino acid residues on the loop (G93-Y111) that interacts with heme. Therefore, these ions seemed to enhance the stability of heme insertion by increasing the loop's steadiness. The basic N-terminal end, together with helix α4 and loop (G126 to Y136), contributed positive charge to the region. In contrast, the acidic C-terminal end provided a negatively charged surface, yielding several electrostatic contact points with partner proteins for electron transfer. These exceptional features of Ma-CytcL, along with the structural information of MDH, led us to hypothesize the need for an adapter protein bridging MDH to CytcL within appropriate proximity for electron transfer. With this knowledge in mind, the methanol oxidation complex reconstitution in vitro could be utilized to produce metabolic intermediates at the industry level.