• ALGAE
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• 제36권4호
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• pp.315-326
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• 2021

Ion channels are membrane protein complexes mediating passive ion flux across the cell membranes. Every organism has a certain set of ion channels that define its physiology. Dinoflagellates are ecologically important microorganisms characterized by effective physiological adaptability, which backs up their massive proliferations that often result in harmful blooms (red tides). In this study, we used a bioinformatics approach to identify homologs of known ion channels that belong to 36 ion channel families. We demonstrated that the versatility of the dinoflagellate physiology is underpinned by a high diversity of ion channels including homologs of animal and plant proteins, as well as channels unique to protists. The analysis of 27 transcriptomes allowed reconstructing a consensus ion channel repertoire (channelome) of dinoflagellates including the members of 31 ion channel families: inwardly-rectifying potassium channels, two-pore domain potassium channels, voltage-gated potassium channels (K_v), tandem K_v, cyclic nucleotide-binding domain-containing channels (CNBD), tandem CNBD, eukaryotic ionotropic glutamate receptors, large-conductance calcium-activated potassium channels, intermediate/small-conductance calcium-activated potassium channels, eukaryotic single-domain voltage-gated cation channels, transient receptor potential channels, two-pore domain calcium channels, four-domain voltage-gated cation channels, cation and anion Cys-loop receptors, small-conductivity mechanosensitive channels, large-conductivity mechanosensitive channels, voltage-gated proton channels, inositole-1,4,5-trisphosphate receptors, slow anion channels, aluminum-activated malate transporters and quick anion channels, mitochondrial calcium uniporters, voltage-dependent anion channels, vesicular chloride channels, ionotropic purinergic receptors, animal volage-insensitive cation channels, channelrhodopsins, bestrophins, voltage-gated chloride channels H⁺/Cl^- exchangers, plant calcium-permeable mechanosensitive channels, and trimeric intracellular cation channels. Overall, dinoflagellates represent cells able to respond to physical and chemical stimuli utilizing a wide range of G-protein coupled receptors- and Ca²⁺-dependent signaling pathways. The applied approach not only shed light on the ion channel set in dinoflagellates, but also provided the information on possible molecular mechanisms underlying vital cellular processes dependent on the ion transport.

• Journal of Microbiology and Biotechnology
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• 제33권7호
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• pp.857-863
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• 2023

Pathogenic bacteria that colonize the human intestinal tract have evolved strategies to overcome acidic conditions when they pass through the gastrointestinal tract. Amino acid-mediated acid resistance systems are effective survival strategies in a stomach that is full of amino acid substrate. The amino acid antiporter, amino acid decarboxylase, and ClC chloride antiporter are all engaged in these systems, and each one plays a role in protecting against or adapting to the acidic environment. The ClC chloride antiporter, a member of the ClC channel family, eliminates negatively charged intracellular chloride ions to avoid inner membrane hyperpolarization as an electrical shunt of the acid resistance system. In this review, we will discuss the structure and function of the prokaryotic ClC chloride antiporter of amino acid-mediated acid resistance system.

• Journal of Periodontal and Implant Science
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• 제45권2호
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• pp.69-75
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• 2015

Purpose: Salivary fluid formation is primarily driven by Ca2+-activated, apical efflux of chloride into the lumen of the salivary acinus. The anoctamin1 protein is an anion channel with properties resembling the endogenous calcium-activated chloride channels. In order to better understand the role of anoctamin proteins in salivary exocrine secretion, the expression of the ten members of the anoctamin gene family in the mouse submandibular gland was studied. Methods: Total RNA extracted from mouse submandibular salivary glands was reverse transcribed using primer pairs to amplify the full-length coding regions of each anoctamin gene and was subcloned into plasmid vectors for DNA sequencing. Alternative splice variants were also screened by polymerase chain reaction using primer pairs that amplified six overlapping regions of the complementary DNA of each anoctamin gene, spanning multiple exons. Results: Multiple anoctamin transcripts were found in the mouse submandibular salivary gland, including full-length transcripts of anoctamin1, anoctamin3, anoctamin4, anoctamin5, anoctamin6, anoctamin9, and anoctamin10. Exon-skipping splicing in the N-terminal exons of the anoctamins1, anoctamin5, and anoctamin6 genes resulted in multiple alternative splice variants. No expression of anoctamin2, anoctamin7, or anoctamin8 was found. Conclusions: The predominant anoctamin transcript expressed in the mouse submandibular gland is anoctamin1ac. The chloride channel protein produced by anoctamin1ac is likely responsible for the $Ca^{2+}$-activated chloride efflux, which is the rate-limiting step in salivary exocrine secretion.

• Journal of Photoscience
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• 제4권1호
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• pp.23-30
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• 1997

Leaf movements in nyctinastic plants are produced by changes in the turgor of extensor and flexor cells, collectively called motor cells, in opposing regions of the leaf movement organ, the pulvinus. In Samanea saman, a tropical tree of the legume family, extensor cells shrink and flexor cells swell to bend the pulvinus and fold the leaf at night, whereas extensor cells swell and flexor cells shrink to straighten the pulvinus and extend the leaf in the daytime. These changes are caused by ion fluxes primarily of potassium and chloride, across the plasma membrane of the motor cells. These ion fluxes are regulated by exogenous light signals and an endogenous biolgical clock. Inward-directed K$^+$ channels are closed in extensor and open in flexor cells in the dark period, while these channels are open in extensor and closed in flexor cells in the light period. Blue light opens the closed K$^+$ channels in extensor and closes the open them in flexor cells during darkness. Illumination of red light followed by darkness induces to open the closed K$^+$ channels in flexor and to close the open K$^+$ channels in extensor cells in the light. The dynamics of K$^+$ channels in motor cells that are controlled by light signals are consistent with the behavior of the pulvini in intact plants. Therefore, these cell types are an attractive model system to elucidate regulations of ion transports and their signal transduction pathways in plants. This review is focused on light-controlled ion movements and regulatory mechanisms involved in phosphoinositide signaling in leaf movements in nyctinastic plants.

• The Korean Journal of Physiology and Pharmacology
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• 제21권4호
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• pp.439-447
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• 2017

Myotonia congenita (MC) is a genetic disease that displays impaired relaxation of skeletal muscle and muscle hypertrophy. This disease is mainly caused by mutations of CLCN1 that encodes human skeletal muscle chloride channel (CLC-1). CLC-1 is a voltage gated chloride channel that activates upon depolarizing potentials and play a major role in stabilization of resting membrane potentials in skeletal muscle. In this study, we report 4 unrelated Korean patients diagnosed with myotonia congenita and their clinical features. Sequence analysis of all coding regions of the patients was performed and mutation, R47W and A298T, was commonly identified. The patients commonly displayed transient muscle weakness and only one patient was diagnosed with autosomal dominant type of myotonia congenita. To investigate the pathological role of the mutation, electrophysiological analysis was also performed in HEK 293 cells transiently expressing homo-or heterodimeric mutant channels. The mutant channels displayed reduced chloride current density and altered channel gating. However, the effect of A298T on channel gating was reduced with the presence of R47W in the same allele. This analysis suggests that impaired CLC-1 channel function can cause myotonia congenita and that R47W has a protective effect on A298T in relation to channel gating. Our results provide clinical features of Korean myotonia congenita patients who have the heterozygous mutation and reveal underlying pathophyological consequences of the mutants by taking electrophysiological approach.

• Bulletin of the Korean Chemical Society
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• 제13권4호
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• pp.395-398
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• 1992

Rate of solvolysis of dansyl chloride in aqueous binary mixtures of acetone, methanol and ethanol are reported. Kinetic solvent isotope effects in methanol and product selectivities in alcohol-water mixtures are also reported. Kinetic data are interpreted with the Grunwald-Winstein and Kivinen equations. The value of $k_{CH3OH}/k_{CH3OD}=1.76$ suggests that a general have catalyzed and/or an $S_AN$ pathway is operative in methanol, a less polar solvent. Rate-rate profiles for solvolysis of dansyl chloride in the aqueous binary media indicate a change in reaction channel from $S_AN$ (in less polar media) to $S_N2$ (in more polar media) mechanism.

• 대한약학회:학술대회논문집
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• 대한약학회 2001년도 Proceedings of the Pharmaceutical Society of Korea
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• pp.124.3-125
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• 2001

• 대한화학회지
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• 제34권3호
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• pp.304-306
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• 1990

• Bulletin of the Korean Chemical Society
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• 제28권4호
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• pp.574-580
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• 2007

Full-length chloride intracellular channel protein 1 (CLIC1) is a member of the family of proteins related to bovine chloride intracellular channel p64. Mutations in the SEDL gene cause spondyloepiphyseal dysplasia tarda (SEDT), a rare X-linked chondrodysplasia. The link between the intracellular chloride channels and SEDL is an important step toward understanding their functional interplay. In the present study, CLIC1 protein was subcloned into the pGEX-KG vector and overexpressed in XL-1 blue cells. We developed a large-scale expression system composed of glutathione S-transferase (GST) fused with a 240-amino-acid CLIC1 protein in Escherichia coli. The soluble CLIC1 protein was successfully purified to homogeneity, and its purity, identity, activity and conformation were determined using SDS-PAGE, MALDI-MS, biophotometer and circular dichroism spectroscopic studies. The binding of both CLIC1 and SEDL proteins in vitro was detected by BIAcore biosensor and fluorescence measurements.

• 한국수정란이식학회지
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• 제19권2호
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• pp.101-112
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• 2004

본 연구는 포유류인 $Cl^-$통로의 존재 여부를 확인하기 위하여 수행되었다. $Cl^-$통로는 비흥분성 세포에서 용적변화와 pH 조절, 이온운반계 등에 중요한 역할을 수행하므로 활발한 세포분화가 이루어지는 난자의 생존과 기능에 필수적 $Cl^-$통로 존재 여부를 확인하고자 하였다. 단일통로 전류를 기록하는 patch clamp 기법을 이용하였다. 세포외(pipette) 용액을 140 mM NaCl로 하고 세포내(bath) 용액을 70 mM, 140 mM, 280 mM NaCl로 바꾸어 주었을 때, 역전압($E_{rev}$) 은 $Cl^-$평형전압을 반영하는 -9.8${\pm}$0.5mV, 0mV, 11.5${\pm}$0.2mV로 변화하였다(n=4). 이런 결과는 이론치와 약간의 편차를 보이고 있으므로 이를 보정하기 위항 Goldman-Hodgkin-Katz(GHK) 식을 이용한 결과 $Cl^-$에 대한 $Na^+$의 투과성 ($P_{Na}$/$P_{Cl}$/)은 0.25로 추측할 수 있었다. Inside out patch mode에서 Pipette 용액과 bath 용액에 imperment인 NMDG-Cl로 구성하였을 경우 단일 통로가 기록되었다.(n=22). 막전압이 증가함에 따라 전도도(conductance)가 같이 증가하였으며 전류-전압 관계는 outward rectification을 보였다. 이런 특성을 나타내는 단일통로로 전류는 선택적 $Cl^-$통로 차단제인 IAA-94(indanyloxyacetic acid 94)에 의해 농도 의존적으로 차단되었으며 가역적으로 회복되었다. IAA의 $IC_{50}$은 32.6$\mu\textrm{M}$이었다. DIDS(4,4'-diisothiocyan ostillben-2-2'disulfonic acid)에 의해서는 전류크기가 감소 되는 빠른 억제기전을 나타내는 차단효과를 보였으며 시간경과에 따라 seblevel로 감소되는 subconductance block 이 나타났다. 이상의 결과는 햄스터 난자에서 chloride 통로가 존재하여, 물질이동과 pH 조절기능을 나타내는 외향성 전류-전압관계를 보이는 $Cl^-$ 통로의 성상으로 미루어 볼 때 난자의 pH 조절과 용적조절과 같은 생리적 환경 조성에 관여할 것으로 추정된다.