• Proceedings of the Korean Biophysical Society Conference
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• 1996.07a
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• pp.34-34
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• 1996

Previously we showed that $Ba^{2+}$ block of large conductance $Ca^{2+}$-activated $K^{+}$ (BK) channel was larger in the planar lipid bilayer formed with negatively-charged phosphatidylserine (PS) than neutral phosphatidylethanolamine (PE). In this work, have studied the blocking effect of two $K^{+}$ channel blockers with different mechanisms of action, $Ba^{2+}$ and tetraethylammonium (TEA), on BK channels of rat skeletal muscle. (omitted)itted)

• Bulletin of the Korean Chemical Society
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• v.31 no.2
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• pp.372-378
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• 2010

Membrane disruption by an antimicrobial peptide, protegrin-1 (PG-1), was investigated by measuring the $^2H$ solid-state nuclear magnetic resonance (SSNMR) spectra of 1-palmitoyl-$d_{31}$-2-oleoyl-sn-glycero-3-phosphatidylcholine (POPC_$d_{31}$) in the mixture of PG-1 and POPC_$d_{31}$ lipids deposited on thin cover-glass plates. The experimental line shapes of anisotropic $^2H$ SSNMR spectra measured at various peptide-to-lipid (P/L) ratios were simulated reasonably by assuming the mosaic spread of bilayers containing pore structures or the coexistence of the mosaic spread of bilayers and a fast-tumbling isotropic phase. Within a few days of incubation in the hydration chamber, the pores were formed by the peptide in the POPC_$d_{31}$ and POPC_$d_{31}$/cholesterol membranes. However, the formation of the pores was not clear in the POPC_$d_{31}$/1-palmitoyl-2-oleoyl-sn-glycero-3-phosphatidylglycerol (POPG) membrane. Over a hundred days after hydration, a rapidly rotating isotropic phase increased in the POPC_$d_{31}$ and the POPC_$d_{31}$/cholesterol membranes with the higher P/L ratios, but no isotropic phase appeared in the POPC_$d_{31}$/POPG membrane. Cholesterol added in the POPC bilayer acted as a stabilizer of the pore structure and suppressed the formation of a fast-tumbling isotropic phase.

• Bulletin of the Korean Chemical Society
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• v.31 no.5
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• pp.1187-1191
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• 2010

$^1H-^{15}N$ heteronuclear dipolar coupling solid-state NMR experiments on lipid bilayer or bicelle samples are very useful for the structural studies of membrane proteins. However, to study these biological samples using solid-state NMR, a specific probe with high efficiency and high capability is required. In this paper, we describe the optimized design, construction, and efficiency of a 400 MHz wide-bore $^1H-^{15}N$ solid-state NMR probe with 5-mm solenoidal rf coil for high power, multi-pulse sequence experiments, such as 2D PISEMA or 2D SAMMY.

• Molecules and Cells
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• v.19 no.3
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• pp.356-364
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• 2005

Intramolecular excimer formation of 1,3-di(1-pyrenyl) propane(Py-3-Py) and fluorescence polarization of 1,6-diphenyl-1,3,5-hexatriene (DPH) were used to evaluate the effect of ethanol on the rate and range of lateral and rotational mobilities of bulk bilayer structures of synaptosomal plasma membrane vesicles (SPMVs) from the bovine cerebral cortex. Ethanol increased the excimer to monomer fluorescence intensity ratio (I'/I) of Py-3-Py in the SPMVs. Selective quenching of both DPH and Py-3-Py by trinitrophenyl groups was used to examine the range of transbilayer asymmetric rotational mobility and the rate and range of transbilayer asymmetric lateral mobility of SPMVs. Ethanol increased the rotational and lateral mobility of the outer monolayer more than of the inner one. Thus ethanol has a selective fluidizing effect within the transbilayer domains of the SPMVs. Radiationless energy transfer from the tryptophans of membrane proteins to Py-3-Py was used to examine both the effect of ethanol on annular lipid fluidity and protein distribution in the SPMVs. Ethanol increased annular lipid fluidity and also caused membrane proteins to cluster. These effects on neuronal membranes may be responsible for some, though not all, of the general anesthetic actions of ethanol.

• Bulletin of the Korean Chemical Society
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• v.34 no.4
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• pp.1120-1126
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• 2013

The syndecan family consists of four transmembrane heparan sulfate proteoglycans present in most cell types and each syndecan shares a common structure containing a heparan sulfate modified extracellular domain, a single transmembrane domain and a C-terminal cytoplasmic domain. To get a better understanding of the mechanism and function of syndecan-4 which is one of the syndecan family, it is crucial to investigate its three-dimensional structure. Unfortunately, it is difficult to prepare the peptide because it is membrane-bound protein that transverses the lipid bilayer of the cell membrane. Here, we optimize the expression, purification, and characterization of transmembrane, cytoplasmic and short extracellular domains of syndecan4 (syndecan-4 eTC). Syndecan-4 eTC was successfully obtained with high purity and yield from the M9 medium. The structural information of syndecan-4 eTC was investigated by MALDI-TOF mass (MS) spectrometry, circular dichroism (CD) spectroscopy, and nuclear magnetic resonance (NMR) spectroscopy. It was confirmed that syndecan-4 eTC had an ${\alpha}$-helical multimeric structure like transmembrane domain of syndecan-4 (syndecan-4 TM) in membrane environments.

• Bulletin of the Korean Chemical Society
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• v.35 no.5
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• pp.1317-1322
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• 2014

Membrane disruption by an antimicrobial peptide (AMP) was investigated by measuring the $^2H$ solid-state nuclear magnetic resonance spectra of 1-palmitoyl-$d_{31}$-2-oleoyl-sn-glycero-3-phosphatidylcholine (POPC_$d_{31}$) in mixtures of POPC_$d_{31}$/cholesterol and either magainin 2 or aurein 3.3 deposited on thin cover-glass plates. The line shapes of the experimental $^2H$ solid-state nuclear magnetic resonance (SSNMR) spectra were best simulated by assuming the coexistence of a mosaic spread of bilayers containing pore structures and a fasttumbling isotropic phase or a hexagonal phase. Within a few days of incubation in a hydration chamber, an isotropic phase and a pore structure were induced by magainin 2, while in case of aurein 3.3 only an isotopic phase was induced in the presence of a bilayer phase. After an incubation period of over 100 days, alignment of the bilayers increased and the amount of the pore structure decreased in case of magainin 2. In contrast with magainin 2, aurein 3.3 induced a hexagonal phase at the peptide-to-lipid ratio of 1/20 and, interestingly, cholesterol was not found in the hexagonal phase induced by aurein 3.3. The experimental results indicate that magainin 2 is more effective in disrupting lipid bilayers containing cholesterol than aurein 3.3.

• BMB Reports
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• v.32 no.6
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• pp.561-566
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• 1999

CA(1-8)-ME(1-12) [CA-ME], composed of cecropin A(1-8) and melittin(1-12), is a synthetic antimicrobial peptide having potent antibacterial and antitumor activities with minimal hemolytic activity. In order to investigate the effects of the flexible hinge sequence, Gly-Ile-Gly, of CA-ME on antibiotic activity, CA-ME and three analogues, CA-ME1, CA-ME2, and CA-ME3, were synthesized. The Gly-Ile-Gly sequence of Ca-ME was deleted in CA-ME1 and replaced with Pro and Gly-Pro-Gly in CA-ME2 and CA-ME3, respectively. CA-ME1 and CA-ME3 showed a significant decrease in antitumor activity and phospholipid vesicle-disrupting ability. However, CA-ME2 showed similar antitumor and vesicle-disrupting activities, as compared with CA-ME. These results suggest that the flexibility or ${\beta}$-turn induced by Gly-Ile-Gly or Pro in the central part of CA-ME may be important in the electrostatic interaction of the N-terminus cationic ${\alpha}$-helical region with the cell membrane surface and the hydrophobic interaction of the C-terminus amphipathic ${\alpha}$-helical region with the hydrophobic acyl chains in the cell membrane. CA-ME3 exhibited lower antitumor and vesicle-disrupting activities than CA-ME and CA-ME2. This result suggests that the excessive ${\beta}$-turn structure caused by the Gly-Pro-Gly sequence in CA-ME3 seems to interrupt ion channel/pore formation in the lipid bilayer. We concluded that the appropriate flexibility or bilayer. We concluded that the appropriate flexibility or ${\beta}$-turn structure provided by the central hinge is responsible for the effective antibiotic activity of the antimicrobial peptides with the helix-hinge-helix structure.

• Journal of the Korean Society of Food Science and Nutrition
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• v.22 no.3
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• pp.323-327
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• 1993

The effect of soyasaponin on the liposomal phospholipid membrane was investigated by differential scanning calorimetry (DSC). Soyasaponins were obtained and the enthalpy changes and the sizes of cooperative unit of the transition were calculated. The thermograms of L-$\alpha$-dimyristoyl phosphatidylcholine (DMPC) incorporated with soyasaponin showed that the phase transition temperature was significantly lowered and the peak was broadened. This was attributed to the possibility that incorporation of soyasaponin into the lipid bilayers reduced the cooperative unit of phospholipid bilayers. These results indicate soyasaponin might have significant effect on the fluidity of biological membrane.

• Journal of the Korean Magnetic Resonance Society
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• v.15 no.1
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• pp.25-39
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• 2011

Syndecan-4 is a transmembrane heparan sulfate proteoglycan, which is a coreceptor with integrins in cell adhesion. To get better understand the mechanism and function of Syndecan-4, it is critical to elucidate the three-dimensional structure of a single transmembrane spanning region of them. Unfortunately, it is hard to prepare the peptide because syndecan-4 is membrane-bound protein that transverse the lipid bilayer of the cell membrane. Generally, the preparation of transmembrane peptide sample is seriously difficult and time-consuming. In fact, high yield production of transmembrane peptides has been limited by experimental adversities of insufficient yields and low solubility of peptide. Here, we demonstrate experimental processes and results to optimize expression, purification, and NMR measurement condition of Syndecan-4 transmembrane peptide.

• Journal of the Korean Applied Science and Technology
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• v.25 no.3
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• pp.313-321
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• 2008

오래전부터, 많은 여성들은 자신들의 젊음을 연장하고, 외모를 더 아름답게 가꾸고, 이를 죽을 때까지 유지하는 것을 바래왔다. 이에 의사와 약사들뿐만 아니라, 많은 화장품 연구자들도 노화와 관련된 기술개발에 총력을 기울여 왔다. 따라서, 이들 연구자들은 노화방지를 위한 새로운 원료를 찾고, 이를 안정화하고, 피부로 전달하는 기술개발에 항상 관심을 쏟아왔다. 뛰어난 노화방지 화장품 개발을 위해서, Ubiquinone의 일종인 ldebenone에 대해 연구하였고, 이를 비수계 피부유사막 기술을 가지고 캡슐화하고 약물전달하는 연구를 진행하였다. 먼저, 편광현미경(PM, Polarized Microscope), X-선 회절분석(XRD, X-ray Diffractions) 및 시차주사열량계 (DSC, Differential Scanning Calorimetry)를 이용하여 Idebenone을 담지한 피부유사막 액정을 비수계 조건에서 구조 및 열적특성을 조사하였다. 그 결과 비수계 조건에서도 규칙적으로 패킹(Packing)된 지질이중층(Lipid bilayer)과 용매의 연속층으로 이루어진 고밀집된 라멜라(Lamella) 구조의 형성유무와 이때의 상거동을 확인할 수 있었다. 결론적으로 높은 극성도로인해 물분자와 접촉하면 불안정해지는 경향이 있는 Idebenone을 비수계 조건에서 각질층(SC, Stratum Corneum)과 구조 및 조성이 유사한 피부유사막을 디자인하여 안정하게 캡슐화 하였다. 이를 적용한 화장품은 모든 보관조건에서 유화입자의 안정성을 유지함을 확인하였고, Idebenone의 활성역가 또한 $40^{\circ}C$에서 6개월 동안 약 90%이상을 유지하는 우수한 결과를 나타냈다.