• Journal of Photoscience
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• 제8권3_4호
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• pp.87-92
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• 2001

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 investigate the effects of parathyroid hormone (PTH) on the bulk bilayer fluidity of the plasma membrane vesicles isolated from cultured osteoblasts (OB-PMV). In a dose-dependent manner, rat PTH-(1-34) [rPTH-(1-34)] increased the excimer to monomer fluorescence intensity ratio (I'/I) of Py-3-Py and decreased the anisotropy (r) of DPH in OB-PMV. This indicates that PTH increased both the lateral and rotational diffusion of the probes in OB-PMY. Selective quenching of DPH fluorescence by trinitrophenyl groups was utilized to examine the transbilayer fluidity asymmetry of OB-PMV. The anisotropy, limiting anisotropy, and order parameter of DPH in the inner monolayer were 0.024, 0.032, and 0.062 greater than calculated for the outer monolayer of OB-PMY. Selective quenching of DPH fluorescence by trinitrophenyl groups was also utilized to examine the transbilayer effects of PTH on the fluidity of OB-PMV. rPTH-(1-34) had a greater fluidizing effect on the outer monolayer as compared to the inner monolayer of OB-PMV. Thus, it has been proven that PTH exhibits a selective rather than nonselective fluidizing effect within transbilayer domains of OB-PMV.

• Archives of Pharmacal Research
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• 제15권2호
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• pp.152-161
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• 1992

Selective quenching of 1, 6-diphenyl-1, 3, 5-hexatriene (DPH) by trinitrophenyl groups was utilized to examine the transbilayer fluidity asymmetry of model membranes of phospholipids (SPMVPL) extracted from synaptosomal plasma membrane vesicles (SPMV). The polarization (P), anisotropy (r), limiting anisotropy $(r_\infty$), and order parameter (S) of DPH in the inner monolayer were 0.019, 0.014, 0.018, and 0.047, respectively, greater than calculated for the outer monolayer of SPMVPL. Selective quenching of DPH by trinitrophenyl groups was also utilized to examine the effects of n-alkanols on the individual monolayer structure of SPMVPL. n-Alkanols fluidized the hydrocarbon region of bulk SPMVPL and the potencies of n-alkanols up to 1-nonanon increased with carbon chain length. It appears that the potencies in bilayer fluidization increase by 1 order of magnitude as the carbon chain length increases by two carbon atoms. The cut-off phenomenon was reached at 1-decanol, where further increase in hydrocarbon length resulted in a decrease in pharmacological activity. The n-alkanols had greater fluidizing effects on the outer monolayer as compared to the inner monolayer of SPMVPL, even though these selective effects tended to become weaker as the carbon chain length increased. Thus, it has been proven that n-alkanols exhibit selective rather than nonselective fludizing effects within transbilayer domains of SPMVPL.

• Molecules and Cells
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• 제19권3호
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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.

• 대한약리학회지
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• 제28권1호
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• pp.103-114
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• 1992

한국산 2년생 소의 신선한 대뇌피질로 부터 synaptosomal plasma membrane vesicles (SPMV)를 분리한 후 이 SPMV로 부터 추출한 총지질 (SPMVTL) 및 총인지질 (SPMVPL)로서 인공세포막을 제제하였다. SPMVTL 및 SPMVPL의 outer monolayer에 trinitrophenyl groups로 공유결합시킴으로써 형광 probe 1,6-diphenyl-1,3,5-hexatriene (DPH)의 outer monolayer 형광을 선택적으로 소광케한 후 SPMVTL 및 SPMVPL의 inner와 outer monolayer 사이의 비대칭적 유동성 존재 유무를 확인하였던 바 inner monolayer에 비하여 outer monolayer가 유동성이 크다는 것을 확인하였으며 SPMVTL에 비하여 SPMVPL의 유동성이 높았다. 뿐만 아니라 barbiturates는 SPMVTL의 유동성에 대하여는 유의성이 있을 만큼의 증감 효과를 나타내지 않았고 SPMVPL의 유동성은 감소시키되 주로 outer monolayer의 유동성을 감소시킨다는 것을 확인하였으며 barbiturates 종류에 따른 SPMVPL의 총유동성 감소의 크기는 pentobarbital > hexobarbital > amobarbital > phenobarbital의 순이었다.

• 대한약리학회지
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• 제28권2호
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• pp.191-199
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• 1992

한국산 2년생 소의 대뇌피질로부터 synaptosomal plasma membrane vesicles(SPMV)를 분리한 후 이 SPMV로부터 추출한 총지질로서 인공세포막(SPMVTL)을 제제하였다. SPMVTL의 outer monolayer에 trinitrophenyl group을 공유결합시킴으로써 SPMVTL이중층에 분포된 형광 probe 1,6-diphenyl-1,3,5-hexatriene(DPH)중 outer monolayer에 분포된 DPH형광만을 소광케 하였다. 형광분석기를 통하여 SPMVTL의 inner monolayer에 비하여 outer monolayer의 유동성이 크다는 것을 확인하였다. n-Alkanols는 SPMVTL지질 이중층(inner+outer monolayers)의 회전확산운동을 증가시키되 탄소수 두개가 증가될 때마다 그 효력이 약10배 가량 증가된다는 것을 알았다. 그러나 1-decanol의 경우에는 1-nonanol에 비하여 그 효력이 낮아지는 소위 cut-off현상이 나타났다. 뿐만 아니라 n-alkanols는 SPMVTL의 inner monolayer에 비하여 outer monolayer의 회전확산운동을 주로 증가시켰다. 하지만 n-alkanols의 탄소수가 증가됨에 따라 outer monolayer에 대한 선택적인 작용이 감소된다는 것도 확인되었다.

• Journal of Photoscience
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• 제12권2호
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• pp.67-73
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• 2005

Fluorescent probe techniques were used to evaluate the effect of dibucaine.HCl on the physical properties (transbilayer asymmetric lateral mobility, annular lipid fluidity and protein distribution) of synaptosomal plasma membrane vesicles (SPMV) isolated from bovine cerebral cortex. An experimental procedure was used based on selective quenching of 1,3-di(l-pyrenyl)propane (Py-3-Py) by trinitrophenyl groups, and radiationless energy transfer from the tryptophans of membrane proteins to Py-3-Py. Dibucaine.HCl increased the bulk lateral mobility, and annular lipid fluidity in SPMV lipid bilayers, and had a greater fluidizing effect on the inner monolayer than the outer monolayer. The magnitude of increasing effect on annular lipid fluidity in SPMV lipid bilayer induced by dibucaine.HCl was significantly far greater than magnitude of increasing effect of the drug on the lateral mobility of bulk SPMV lipid bilayer. It also caused membrane proteins to cluster. These effects of dibucaine.HCl on neuronal membranes may be responsible for some, though not all, of the local anesthetic actions of dibucaine.HCl.

• BMB Reports
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• 제33권6호
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• pp.541-547
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• 2000

Fluorescence polarization of 1,6-diphenyl-1,3,5-hexatriene (DPH) was used to evaluate the effects of chlorpromazine HCl on the range of the rotational mobility of bulk bilayer structure of the synaptosomal plasma membrane vesicles (SPMV) isolated from a bovine brain. In a dose-dependent manner, chlorpromazine HCl increased the anisotropy (r), limiting anisotropy ($r_{\infty}$) and order parameter (S) of DPH in the membranes. Cationic 1-[4-(trimethylammonio)-phenyl]-6-phenylhexa-1,3,5-hexatriene (TMA-DPH) and anionic 3-[p-(6-phenyl)-1,3,5-hexatrienyl]-phenylpropionic acid (PRO-DPH) were utilized to examine the range of transbilayer asymmetric rotational mobility of the neuronal membranes. The anisotropy (r) of TMA-DPH in the inner monolayer was 0.034 greater than the value of PRO-DPH in the outer monolayer of the membranes. Both cationic TMA-DPH and anionic PRO-DPH were also used to examine the transbilayer asymmetric effects of chlorpromazine HCl on the range of rotational mobility of the membranes. Chlorpromazine HCl have a decreasing effects on the rotational mobility of the bulk bilayer structures and have a greater decreasing effect on the mobility of the inner monolayer as compared to the outer monolayer of the membranes. It has been proven that chlorpromazine HCl exhibit a selective rather than nonselective fluidizing effect within the transbilayer domains of the SPMV.

• BMB Reports
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• 제37권5호
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• pp.603-611
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• 2004

Fluorescence probes located in different membrane regions were used to evaluate the effects of chlorpromazine HCl on structural parameters (transbilayer lateral mobility, annular lipid fluidity, protein distribution, and lipid bilayer thickness) of synaptosomal plasma membrane vesicles (SPMVs) isolated from bovine cerebral cortex. The experimental procedure was based on the selective quenching of 1,3-di(1-pyrenyl)propane (Py-3-Py) by trinitrophenyl groups, radiationless energy transfer from the tryptophan of membrane proteins to Py-3-Py, and energy transfer from Py-3-Py monomers to 1-anilinonaphthalene-8-sulfonic acid (ANS). In this study, chlorpromazine HCl decreased the lateral mobility of Py-3-Py in a concentration dependent-manner, showed a greater ordering effect on the inner monolayer than on the outer monolayer, decreased annular lipid fluidity in a dose dependent-manner, and contracted the membrane lipid bilayer. Furthermore, the drug was found to have a clustering effect on membrane proteins.

• Archives of Pharmacal Research
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• 제27권6호
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• pp.653-661
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• 2004

Fluorescence probes located in different membrane regions were used to evaluate the effect of dopamine$.$HCI on the structural parameters (transbilayer lateral mobility, annular lipid fluidity, protein distribution, and thickness of the lipid bilayer) of synaptosomal plasma membrane vesicles (SPMV), which were obtained from the bovine cerebral cortex. An experimental procedure was used based on selective quenching of 1,3-di(1-pyrenyl)propane (Py-3-Py) by trinitrophenyl groups, and radiationless energy transfer from the tryptophan of membrane pro-teins to Py-3-Py and energy transfer from Py-3-Py monomers to 1-anilinonaphthalene-8-sulfonic acid (ANS) was also utilized. Dopamine$.$HCI increased both the bulk lateral mobility and annular lipid fluidity, and it had a greater fluidizing effect on the inner monolayer than on the outer monolayer. Furthermore, the drug had a clustering effect on membrane proteins.

• 대한약리학회지
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• 제29권1호
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• pp.149-156
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• 1993

Ethanol이 암세포 증식 속도에 미치는 직접적인 영향 검색의 일환으로 배양된 mouse myeloma cell line Sp2/0-Ag14로부터 분리한 형질막 (Sp2/0-PMV) 유동성에 미치는 ethanol의 영향을 형광분석법으로 측정하였다. 그 결과 ethanol은 Sp2/0-PMV 지질 이중층 측방확산운동의 범위와 속도를 증가시켰고 회전확산운동 범위도 증가시켰다. 특히 ethanol은 Sp2/0-PMV 지질이중층 중 내부단층 (inner monolayer)에 비하여 비교적 선택적으로 외부단층 (outer monolayer)의 회전확산운동 범위를 증가시킨다는 것을 확인하였다.