• Bulletin of the Korean Chemical Society
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• v.27 no.1
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• pp.87-92
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• 2006

Fas-associated death domain protein (FADD) recruits and activates procaspase-8 through interactions between the death effector domains of these two proteins. Cellular FLICE-inhibitory protein (c-FLIP) was identified as a molecule with sequence homology to caspase-8. It has been postulated that c-FLIP prevents formation of the competent death-inducing signaling complex in a ligand-dependent manner, through its interaction with FADD and/or caspase-8. However, the interaction of FADD and $c-FLIP_s$ (short form) in apoptosis signaling has been controversially discussed. We show the purification and the characterization of human full-length FADD and $c-FLIP_s$ expressed in Escherichia coli. The purified FADD and $c-FLIP_s$ are shown as homogeneity, respectively, in SDS-PAGE analysis and light-scattering measurements. The folding properties of the $\alpha$-helical structure of FADD and the super-secondary structure of $c-FLIP_s$ proteins were characterized by circular dichroism spectroscopy. Furthermore, we report here a series of biochemical and biophysical data for FADD-$c-FLIP_s$ binding in vitro. The binding of both FADD and $c-FLIP_s$ proteins was detected by BIAcore biosensor, fluorescence measurement, and size-exclusion column (SEC).

• Korean Journal Plant Pathology
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• v.1 no.3
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• pp.199-206
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• 1985

Viroids are the smallest. well-characterized infectious agents presently known. and so far viroids have been found only in higher plants. The structures of viroid-molecules are single-stranded, covalently closed circular RNA molecules with a range of 240 to 380 nucleotides according to the various viroids. Viroids are remarkable not only as a new category of pathogen, which cause economically important diseases, but also as an excellent model system for biochemical and biophysical investigations because of their small size, relative stability and their self-replication. Four different patato spindle tuber viroid isolates, which express the different symptoms on the same host plant exchange only 2 to 6 nucleotides in the total number of 359 nucleotides, but now the mechanism of viroid pathogenicity is not explained fully. Viroid-melecules are replicated without any special viroid-associated proteins, and during the process of viroid replication oligomeric viroid-associated RNAs are detected at nuclei of viroid infected leaf tissue. The mechanism of viroid replication can now be illustrated according to a possible explanation of rolling-circle system. Although the rapid progress have been made in elucidation of the biochemical and biophysical properties of PSTV and other viroids, the mechanism of viroid replication and pathogenicity is less known and is still a matter of speculation. When these problems can be sufficiently explained, the viroid molecule could play an important role as an available vector in plant genetic engineering.

• Proceedings of the Korean Biophysical Society Conference
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• 2003.06a
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• pp.23-23
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• 2003

It has been suggested that the impairment of smooth muscle cell (SMC) function by alterations in the $Ca^{2+}$-activated $K^{+}$ ( $K_{Ca}$ ) channels accounts for the reduction in coronary reserve during left ventricular hypertrophy (LVH). However, this hypothesis has not been fully investigated. The main goal of this study was to assess whether the properties of $K_{Ca}$ channels in coronary SMCs were altered during LVH. New Zealand white rabbits (0.8-1.0 kg) and Sprague-Dawley rats (300-400 g) were randomly selected to receive either an injection of isoproterenol (300 $\mu\textrm{g}$/kg body weight) or an equal volume of 0.9％ saline (1 mL/kg body weight). The animals developed LVH 10 days after injection. In patch-clamp experiments, the unitary current amplitude and open probability for the $K_{Ca}$ channels were significantly reduced in LVH patches compared with control patches. The concentration-response curve of the $K_{Ca}$ channel to [C $a^{2+}$]$_{i}$ was shifted to the right. Inhibition of the $K_{Ca}$ channels with TEA was more pronounced in LVH cells than in the control cells. The whole-cell currents of $K_{Ca}$ channels were reduced during LVH. Western blot analysis indicated no differences in $K_{Ca}$ channel expression between the control and LVH coronary SM membranes. In contraction experiments, the effect of a high $K^{+}$concentration on the resting tension of the LVH coronary artery was greater than on that of the control. The effect of TEA on the resting tension of the LVH coronary artery was reduced as compared with the effect on the control. Our findings imply a novel mechanism for reduced coronary reserve during LVH.ing LVH.

• Proceedings of the Korean Biophysical Society Conference
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• 2003.06a
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• pp.39-39
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• 2003

In our previous study, WEHI-231, an immature B cell line, showed intractable increase in [C $a^{2+}$]$_{c}$ after the B-cell receptor (BCR) ligation and treatment with 2-aminoethoxydiphenylborate (2-APB), which was never observed in Bal-17, a mature B cell line (Nam et al., 2003, FEBS Lett). In this study, a whole cell voltage clamp study revealed a specific expression of a novel type of $K^{+}$ current, namely voltage-independent background-type $K^{+}$ channels (IK-bg), in WEHI-231 cells. IK-bg was dramatically increase by the application of 2-APB (50 $\square$M), which induced severe hyperpolarization of WEHI-231 from -45 ㎷ to -90 ㎷, When dialyzed with $Mg^{2+}$ and ATP-free pipette solution, a spontaneous development of IK-bg and membrane hyperpolarization were observed. IK-bg was insensitive to classical $K^{+}$ channel blockers (TEA, glibenclamide, $Ba^{2+}$(1 mM)), whereas blocked by quinine and quinidine in a voltage-dependent manner ($IC_{50}$/=6~9 $\square$M at +60㎷). Phorbol myrstate, a PKC activator, decreased the amplitude of IK-bg. Extracellular acidification (pH 6.5) slightly inhibited IK-bg. Arachidonic acid, riluzole, or hyposmotic stress could not affect the IK-bg after the full development by the intracellular dialysis with Mg-ATP-free solution. In a cell-attached mode of single channel recording from WEHI231, we found two types of voltage-independent $K^{+}$ channels with unitary conductance of 300 pS and 120 pS, respectively. Both channels showed very short mean open times and their open probabilities were increase by the application of 2-APB. In Bal-17 cells, no such $K^{+}$ current was observed in 50 cells tested. In summary, WEHI-231 immature B cells express background $K^{+}$ channels. The pharmacological properties and the large unitary conductance suggest that novel types of two-pore domain $K^{+}$ channels (2-P-K channels) might be expressed in WEHI-231, which may provide an intriguing targets of signal transduction in the immature B lymphocytes.e B lymphocytes.

• Journal of Life Science
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• v.17 no.11
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• pp.1517-1522
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• 2007

Gap junction channels formed by two adjacent cells allow the passage of small molecules up to ${\sim}\;1\;kDa$ between them. Hemichannel (connexon or half of gap junction) also behaves as a membrane channel like sodium or potassium channels in a single cell membrane. Among 26 types of connexin (Cx), $Cx32^*43E1$ (a chimera in which the first extracellular loop of Cx32 has been replaced with that of Cx43), Cx38, Cx46, and Cx50 form functional hemichannels as well as gap junction channels. Although it is known that Xenopus oocytes express endogenous connexin 38 (Cx38), its biophysical characteristics at single channel level are poorly understood. In this study, we performed single channel recordings from single Xenopus oocytes to acquire the biophysical properties of Cx38 including voltage-dependent gating and permeation (conductance and selectivity). The voltage-dependent fast and slow gatings of Cx38 hemichannel are distinct. Fast gating events occur at positive potentials and their open probabilities are low. In contrast, slow gatings dominate at negative potentials with high open probabilites. Based on hi-ionic experiments, Cx38 hemichannel is anion-selective. It will be interesting to test whether charged amino acid residues in the amino terminus of Cx38 are responsible for voltage gatings and permeation.

• Journal of the Korean Chemical Society
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• v.55 no.3
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• pp.335-340
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• 2011

The solution properties of sodium n-dodecyl sulfate, as an anionic surfactant in the presence of a cationic watersoluble 5, 10, 15, 20-tetrakis (N-methylpyridinium-4-yl) porphyrin (TMPyP) has been comprehensively studied by means of conductometry, UV-vis and resonance light scattering (RLS) spectroscopies. The results represent the decreasing of critical micelle concentration of SDS solution due to increasing of TMPyP concentration. The stabilization of SDS micelle is due to neutralization of negative charge at the micelle surface. The presence of three different species of TMPyP in SDS solution has been unequivocally demonstrated: free porphyrin monomers, porphyrin monomers or aggregates bound to the micelles, and nonmicellar porphyrin/surfactant aggregates. Our results show SDS induced an aggregation in TMPyP. In fact two kinds of J-aggregations were observed: one of them for porphyrin monomers or aggregates bound to the micelles and the other for nonmicellar porphyrin/surfactant aggregates. However, the results represent the electrostatic interaction of TMPyP with SDS anion below the cmc.

• BMB Reports
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• v.42 no.6
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• pp.373-379
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• 2009

5-Hydroxyuridine (5-OHU) is a major lesion of uridine and cytosine produced in RNA by various chemical oxidants. To elucidate its biochemical and biophysical effects on RNA replication, the site-specifically modified oligoribonucleotides containing 5-OHU were synthesized with C5-hydroxy-5'-ODMTr-2'-TBDMS-uridine phosphoramidite using automated solid phase synthesis. The base-pairing properties of nucleotides opposite 5-OHU in 24 mer oligoribonulcleotides with dNTP were studied using three reverse transcriptases (Super-$Script^{TM}II$-, AMV-, MMLV-RT) in cDNA synthesis. Adenine as well as guanine was incorporated preferentially by all reverse transcriptases. In the UV-melting temperature experiment, the results from the relative stabilities of the base pairs were A : 5-OHU > G : 5-OHU > T : 5-OHU $\approx$ C : 5-OHU. Circular Dichroism (CD) studies showed that DNA-RNA containing 5- OHU heteroduplexes exhibit a similar conformation between the A-type RNA and B-type DNA. These results suggest that 5- OHU from oxidative damage was mainly influenced by adenine mismatch.

• Korean Journal of Environment and Ecology
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• v.19 no.3
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• pp.215-230
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• 2005

To analyze physical and ecological characteristics of sand streams, Cheongmi, Yanghwa and Bokha streams on the similar environment were Lnvestigated and estimated. According to the estimation results of physical structures, ecological properties of the Cheogmi and Yanghwa streams were classified as 'fair to good', and the Bokha stream was classified as 'fair'; commonly, the representing streams were comparatively active on meandering and accumulation; on the other hand, erosion was less active. As a result of the study for flora on this 3 streams, $Phaiaris\;arundinacfa\rightarrowPhragmites\;japonica\rightarrowPhragmites\;communis\rightarrowSalix\;koreensis\rightarrowFraxinus\;rhynchophylla\rightarrowRobinia\;pseudo-acacia-Acer$ ginnala communities were developed in order of being closest to the water shore. In case of benthic macroinvertebrates, Ephemera striga ta Eaton and Cajopteryx atrata Selys, usually inhabited in the uncontaminated water, were investigated as common species. Besides, Limnodrilus gotoi Hatai and Family Chironomidae sp. 1 were the common dominant benthic macroinvertebrates.

• Journal of Applied Biological Chemistry
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• v.52 no.3
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• pp.93-102
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• 2009

Polyunsaturated fatty acids (PUFA), especially eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) have significantly beneficial effects on health in relation to cardiovascular, immune, and inflammatory conditions and they are involved in determining the biophysical properties of membranes as well as being precursors for signaling molecules. PUFA biosynthesis is catalyzed by sequential desaturation and fatty acyl elongation reactions. This aerobic biosynthetic pathway was thought to be taxonomically conserved, but an alternative anaerobic pathway for the biosynthesis of PUFA is now known to contain analogous polyketide synthases (PKS). Certain fish oil can be a rich source of PUFA although processed marine oil is generally undesirable as food ingredients because of the associated objectionable flavors that are difficult and cost-prohibitive to remove. Oil-seed plants contain only the 18-carbon polyunsaturated fatty acid alpha-linolenic acid, which is not converted in the human body to EPA and DHA. It is now possible to engineer common oilseeds which can produce EPA and DHA and this has been the focus of a number of academic and industrial research groups. Recent advances and future prospects in the production of EPA and DHA in oilseed crops are discussed here.

• Journal of Microbiology and Biotechnology
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• v.25 no.8
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• pp.1371-1379
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• 2015

The Ca_v1.2 Ca²⁺ channel is essential for cardiac and smooth muscle contractility and many physiological functions. We mutated single, double, and quadruple sites of the four potential Asn (N)-glycosylation sites in the rabbit Ca_v1.2 into Gln (Q) to explore the effects of Nglycosylation. When a single mutant (N124Q, N299Q, N1359Q, or N1410Q) or Ca_v1.2/WT was expressed in Xenopus oocytes, the biophysical properties of single mutants were not significantly different from Ca_v1.2/WT. In comparison, the double mutant N124,299Q showed a positive shift in voltage-dependent gating. Furthermore, the quadruple mutant (QM; N124,299,1359,1410Q) showed a positive shift in voltage-dependent gating as well as a reduction of current. We tagged EGFP to the QM, double mutants, and Ca_v1.2/WT to chase the mechanisms underlying the reduced currents of QM. The surface fluorescence intensity of QM was weaker than that of Ca_v1.2/WT, suggesting that the reduced current of QM arises from its lower surface expression than Ca_v1.2/WT. Tunicamycin treatment of oocytes expressing Ca_v1.2/WT mimicked the effects of the quadruple mutations. These findings suggest that Nglycosylation contributes to the surface expression and voltage-dependent gating of Ca_v1.2.