• Molecules and Cells
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• 제37권8호
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• pp.592-597
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• 2014

The cell membrane provides critical cellular functions that rely on its elaborate structure and organization. The structure of turtle membranes is an important part of an ongoing study of erythrocyte membranes. Using a combination of atomic force microscopy and single-molecule force spectroscopy, we characterized the turtle erythrocyte membrane structure with molecular resolution in a quasi-native state. High-resolution images both leaflets of turtle erythrocyte membranes revealed a smooth outer membrane leaflet and a protein covered inner membrane leaflet. This asymmetry was verified by single-molecule force spectroscopy, which detects numerous exposed amino groups of membrane proteins in the inner membrane leaflet but much fewer in the outer leaflet. The asymmetric membrane structure of turtle erythrocytes is consistent with the semi-mosaic model of human, chicken and fish erythrocyte membrane structure, making the semi-mosaic model more widely applicable. From the perspective of biological evolution, this result may support the universality of the semi-mosaic model.

• 대한의생명과학회지
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• 제11권1호
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• pp.57-61
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• 2005

The number of sites at which a protein can be readily cleaved by a proteolytic enzyme is greatly influenced by its three-dimensional structure. For native, properly-folded proteins both the rate of cleavage and number of sites at which cleavage takes place are usually much less than for the denatured protein. In order to compare the tertiary structure of recombinant HepG2 type glucose transporter with that of its native counterpart in the erythrocyte, the pattern of tryptic cleavage of the protein expressed in insect cell membranes was therefore examined. After 30 minutes digestion, a fragment of approximate Mr 19,000-21,000 was generated. In addition to this, there were two less intensely stained fragments of apparent Mr 28,000 and 17,000. The pattern of labelling was similar up to 2 hours of digestion. However, the fragments of Mr 19,000-21,000 and Mr 17,000 were no longer detectable after 4 hours digestion. The observation of a very similar pattern of fragments yielded by tryptic digestion of the HepG2 type transporter expressed in insect cells suggests that the recombinant protein exhibits a tertiary structure similar if not identical to that of its human counterpart. Also, the endogenous sugar transporter(s) present in Sf21 cells did not bind cytochalasin B, the potent transporter inhibitor. Therefore, the baculovirus/Spodoptera frugiperda (Sf) cell expression system could be very useful for production of large amounts of human glucose transporters, heterologously.

• 대한의생명과학회지
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• 제8권4호
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• pp.211-215
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• 2002

The baculovirus/Sf9 cell expression can be employed as a powerful system for producing large amounts of the human erythrocyte glucose transporter, GLUT1 heterologously In order to exploit the system further, it is necessary to develop a convenient method for demonstrating that the transporter expressed in insect cells is biologically active. To achieve this, we have expressed the human CLUT1 in insect cells and photolabelled the expressed protein with [$^3$H] cytochalasin B, a potent inhibitor of the human erythrocyte glucose transporter. Subsequently, the labelled proteins were analysed by SDS-polyacrylamide gel electrophoresis (SDS-PAGE). Membranes labelled with [$^3$H] cytochalasln B in the presence of L-Glucose yielded a single sharp peak of labelling of apparent $M_r$ 45,000 on SDS/polyacrylamide gels. The mobility of this peak corresponded exactly to that of the band detected by anti-glucose transporter antibodies on Western blots of membranes prepared from insect cells infected with recombinant virus. In addition, the sharpness of the radioactive peak provides further evidence for the conclusion that the expressed protein is much less heavily and heterogeneously glycosylated than its erythrocyte counterpart. No peak of labelling was seen with the membranes prepared from non-infected Sf9 cells. Furthermore, the incorporation of label into this peak was completely inhibited by the presence of 500 mM-D-Glucose during tile photolabelling procedure, showing the stereoselectivity of the labelling. These evidences clearly show that human glucose transporter expressed in insect cells exhibits native-like biological activity, and that photolabelling with [$^3$H] cytochalasin B can be a convenient means for analysing the biological activity of the transport protein expressed in insect cells.

• The Korean Journal of Physiology and Pharmacology
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• 제4권3호
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• pp.243-251
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• 2000

In order to provide a basis for studying the molecular mechanism of pharmacological action of local anesthetics and to develop a fluorescence spectroscopic method which can detect the microviscosity of native and model membranes using intramolecular excimerization of 1,3-di(l-pyrenyl)propane (Py-3-Py), we examined the effect of lidocaine HCl on the microviscosity of model membranes of phosphatidylcholine fraction extracted from synaptosomal plasma membrane vesicles (SPMVPC). The excimer to monomer fluorescence intensity ratio (I'/I) of Py-3-Py in liquid paraffin was a simple linear function of $T/{\eta}.$ Based on this calibration curve, the microviscosity values of the direct probe environment in SPMVPC model membranes ranged from $234.97{\pm}48.85$ cP at $4^{\circ}C$ to %19.21{\pm}1.11$ cP at $45^{\circ}C.$ At $37^{\circ}C,$ a value of $27.25{\pm}0.44$ cP was obtained. The lidocaine HCl decreased the microviscosity of SPMVPC model membranes in a concentration-dependent manner, with a significant decrease in microviscosity value by injecting the local anesthetic even at the concentration of 0.5 mM. These results indicate that the direct environment of Py-3-Py in the SPMVPC model membranes is significantly fluidized by the lidocaine HCl. Also, the present study explicitly shows that an interaction between local anesthetics and membrane lipids is of importance in the molecular mechanism of pharmacological action of lidocaine HCl.

• International Journal of Oral Biology
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• 제42권4호
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• pp.175-181
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• 2017

The aim of this study was to provide a basis for the molecular mechanism underlying the pharmacological action of ethanol. We studied the effects of 1-propanol on the location of n-(9-anthroyloxy)palmitic acid or stearic acid (n-AS) within the phospholipids of synaptosomal plasma membrane vesicles (SPMV). The SPMV were isolated from the bovine cerebral cortex and liposomes of total lipids (SPMVTL) and phospholipids (SPMVPL). 1-Propanol increased the rotational mobility of inner hydrocarbons, while decreasing the mobility of membrane interface, in native and model membranes. The degree of rotational mobility varied with the number of carbon atoms at positions 16, 12, 9, 6 and 2 in the aliphatic chain of phospholipids in the neuronal and model membranes. The sensitivity of increasing or decreasing rotational mobility of hydrocarbon interior or surface by 1-propanol varied with the neuronal and model membranes in the following order: SPMV, SPMVPL and SPMVTL.

• 한국발생생물학회지:발생과생식
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• 제21권3호
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• pp.287-295
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• 2017

This study was conducted to observe egg and larvae morphological development of carp to obtain basic data for resource conservation and taxonomic research. Brood carp used in the research (total length 67.3-75.5 cm, average $71.0{\pm}3.45cm$) were bred in a circular rearing aquarium ($600{\times}300{\times}100cm$) using a running water system from January to July, 2015. Breeding water temperature was maintained at $23.0-25.0^{\circ}C$(average $24.0^{\circ}C$). Fertilized carp eggs were translucent and globular, and their size was 1.75-1.89 mm (average $1.82{\pm}0.06mm$). Blastoderms formed 10 min after fertilization and reached the two-cell stage 30 min after fertilization. Then, the embryo turned dark and exhibited melanophores, and blood started flowing from the heart across the egg yolk at 42 hrs and 50 min after fertilization. Hatching began 70 hrs and 26 min after fertilization larvae emerged through the egg membrane, starting from the head. The length of prelarvae immediately after hatching was 5.23-5.38 mm (average $5.31{\pm}0.11mm$) the mouth and anus were closed, and the pectoral fin was formed. Postlarvae at 18 days after hatching had a total length of 11.9-13.9 mm (average $12.9{\pm}1.40mm$), separate anal fin and back membranes, and fin ray. Juveniles fish at 35 days after hatching had a total length of 29.9-30.2 mm (average $30.1{\pm}0.13mm$), with the body covered with scales, and the same number of fin rays, color, and shape as their broodstork.

• 대한수의학회지
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• 제34권4호
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• pp.715-725
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• 1994

The ultrastructural investigations of the parotid gland of Korean native goat were carried out by transmission electron microscopy. The results were as follows; 1. The acini of parotid gland were composed of light and dark acinar cells. 2. In the light acinar cells, the secretory granules were classified into three types according to their electron densities and dense bodies. One type of granules was low electron density and had no dense bodies. Another type was low electron density and had dense bodies, and the other type was low electron density and had granular dense bodies. 3. The secretory granules of dark acinar cells showed high electron density and were also calssified into three types by dense bodies as the same way as in the light acinar cells. 4. The intercalated ducts consisted of simple cuboidal epithelium. The nuclei of epithelial cells were oval or round form, located at the central part, and had infolding nuclear membranes and one or two nucleoli. 5. The cells of both of the striated and excretory ducts were composed of light cells, dark cells, specific light cells and basal cells. 6. The nerve terminals were distinguished into two types. One had large granular synaptic vesicles, and another had small agranular synaptic vesicles.

• 대한화학회지
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• 제42권2호
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• pp.143-149
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• 1998

마취제를 purple membrane내 bacteriorhodopsin(bR)에 가했을 때 570nm 흡수 띠는 480nm로 이동한다. 마취제에 의해 유도된 분광학적 변화는 가역적이다. 이러한 평형의 겉보기 pKa(6.3)는 bR에 분산된 마취제의 성질에 의존한다. bR의 전기배향 측정은 비교적 작은 전기장에서도 쉽게 달성될 수 있었으며 이때의 배향각도는 60°이었다. 그러나 마취제로 처리한 bR은 이러한 변화를 관측할 수가 없다. 이들 결과들은 마취제에 의해 발색단과 발색단 주변의 단백질 구조의 민감한 변화가 단백질 매트릭스의 전하를 띈 잔기들의 공간적 배향에 영향을 준 것으로 보인다.

• 한국가축번식학회지
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• 제16권3호
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• pp.247-260
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• 1992

This study was carried out to investigate the histomorphological changes of ovaries obtained from 100 of 1-year-old female Korean loach(misgurnus anguillicaudatus). The light microscopic and ultrastructural changes ofooplasm and follicular membranes of oocytes, were observed by lightand transmission electron microscope during the reproductive cycle. All data were collected from November in 1991 to May in 1992. The results obtained in this study were as follows: The size of the nucleoli and number of the yolk granules increased as the oocytes grown. Yolk granules were loosely deposited in the oocytes as crystalline granules. Due to the presence of large early and late maturing oocytes, their ovaries were enlarged, transparent, granular and yellowish in color. The lattice was broken down at hydration, leaving the egg transparent. As the percentages of fish in LMO and RO stage increased from March to April, mean gonadosomatic index(GSI) values(18.49%) increased. Zona radiata change a squamous into cuboid shape in EMO stage. Processes from the granulosa cells and from the oocyte, microvilli grow and make contact with other in the pore canals of the zona radiata during vitellogenesis, but are withdrawn as the zona radiata becomes more compact and devoid of pore canals during oocyte maturation. Seasonal changes in the microscopic appearance of the ovaries were well correlated with those in both GSI and macroscopic appearance.

• The Korean Journal of Pain
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• 제6권2호
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• pp.237-246
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• 1993

세포막에서 마취제의 작용점을 규명하기 위하여, 마취제의 많은 부분을 차지하는 n-Alkanol을 이용하여, 소의 synaptosomal plasma membrane vesicles(SPMV)에서 n-Alkanol의 침투 정도를 형광 probe를 이용한 형광소광법을 통하여 검색하였다. n-Alkanols는 SPMV 외부 단층(outer monolayer)의 표면에 주로 분포하되 그 탄소수에 비례하여 소수성 부위에 분포되는 양이 증가되는 경향을 나타내었다(1-decanol은 제외). Methanol, Ethanol, 1-propanol, 1-butanol, 1-pentano, 1-hexanol, 1-heptanol, 1-octanol, 1-nonanol 및 1-decanil은 SPMV 외부 단층의 표면(친수성 부위)에 분포되는 것에 비하여 각각 949, 416.8, 214.8, 90.3, 53.7, 15.20, 6.80, 2.00, 1.03 및 2.40 배가 된다는 것을 확인하였다. 1-decanol은 $C_{10}$인데도 불구하고 $C_8$인 1-octanol에 비하여 적은 양이 소수성 부위에 침투 분포한다는 것이 확인되었다.