• 한국미생물학회:학술대회논문집
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• 한국미생물학회 2001년도 추계학술대회
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• pp.97-101
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• 2001

The fusion between viral envelope and target cell membrane is a central step of viral infection, and the fusion proteins located at viral envelope mediate such process. Gp41 of HIV is one of the fusion proteins whose structure and mechanism of membrane fusion had been extensively studied. Functionally important motives of gp41 are the N-terminus fusion peptide, the coiled-coil and the membrane proximal C-peptide regions. The role of these regions during the fusion process had been thoroughly examined. Specially, insertion of the fusion peptide into membrane and conformational change of the coiled-coil and C-peptide regions are assumed to be critical for the fusion mechanism. In addition, the coiled-coil region has been shown to interact with membrane, and the C-peptide region regulates the interaction in a dose dependent manner. Furthermore, fusion defective mutations of the coiled-coil region dramatically changed its binding affinity to membrane. These results suggested that the membrane binding property of the coiled-coil region is important for the fusion activity of gp41, and such property could be modulated by the interaction with the C-peptide region.

• Bulletin of the Korean Chemical Society
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• 제28권10호
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• pp.1756-1762
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• 2007

Envelope proteins of virus contain a segment of hydrophobic amino acids, called as fusion peptide, which triggers membrane fusion by insertion into membrane and perturbation of lipid bilayer structure. Potential fusion peptide sequences have been identified in the middle of L or M proteins or at the N-terminus of S protein in the envelope of human hepatitis B virus (HBV). Two 16-mer peptides representing the N-terminal fusion peptide of the S protein and the internal fusion peptide in L protein were synthesized, and their membrane disrupting activities were characterized. The internal fusion peptide in L protein showed higher activity of liposome leakage and hemolysis of human red blood cells than the N-terminal fusion peptide of S protein. Also, the membrane disrupting activity of the extracellular domain of L protein significantly increased when the internal fusion peptide region was exposed to N-terminus by the treatment of V8 protease. These results indicate that the internal fusion peptide region of L protein could activate membrane fusion when it is exposed by proteolysis.

• Journal of Microbiology
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• 제34권1호
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• pp.7-14
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• 1996

Teh baculovirus gp64 glycoprotein is a major component of the envelope of budded virus (BV) and has been shown that it plays an essential role in the infection process, especially virus-cell membrane fusion. We have cloned Autographa californica Nuclear Polyhedrosis Virus (AcNPV) gp64 protein were examined for membrane fusion activity by using a synchtium formation assay under various conditions. The optimal conditions required for inducing membrane fusion are 1) form pH 4.0 to 4.8 2) 15 min exposure of cells to acidic pH 3) at least 1 .mu.g of gp64 cloned plasmid DNA per 3 * 10$^{6}$ cells 4) and an exposure of cells to acidic pH at 72 h post-transfection. In order to investigate the role of hydrophobicity of the gp64 glycoprotein for the membrane fusion, the two leucine residues (amino acid position at 229 and 230) within hydrophobic region I were substituted to alanine by PCR-derived site-directed mutagenisis and the membrane fusion activity of the mutant was anlaysed. The gp64 glycoprotein carrying double alamine substitution mutation showed no significant difference in fusion activity. This result suggested that minor changes in hydrophobicity at the amino acid position 229 and 230 does not affect the acid-induced membrane fusion activity of the gp64 glycoprotein.

• 한국생물물리학회:학술대회논문집
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• 한국생물물리학회 2003년도 정기총회 및 학술발표회
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• pp.58-58
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• 2003

The envelope glycoprotein of HIV, gp41, mediates the membrane fusion with human cells. The extracellular domain of gp41 has two helical regions. The N-terminus helical region (N-helix) forms trimeric coiled coil, interacts with the C-terminus helical region (C-helix) of gp41 to form a stable helical bundle structure. In this study, we have shown that the N-helix of gp41 has membrane interacting and disrupting abilities. It was localized into the interface of the lipidic phase and head group of the membrane. In contrast, the N-helix region with membrane fusion defective mutations could not bind to membrane. In addition, the N-helix bound on the membrane was released from the membrane by the C-helix, and the complex of the N- and C-helix did not interact with membrane. These results suggested that the membrane binding ability of the N-helix is necessary for the fusion activity of gp41, and such property is possibly controlled by the C-helm.

• 미생물학회지
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• 제24권3호
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• pp.197-203
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• 1986

Streptomyces lavendulae에서 생성된 원형질체 및 원형질체 융합과정에 대한 미세구조 및 행태학적 관찰을 투과 전자현미경과 주사전자현미경을 사용하여 행하였다. 생성된 원행질체들은 고장액에서 대체로 안정하였으나 간혹 변형된 원형질체들이 관찰되었다. 원형질체 융합은 접촉지역 융합지역, 분리지역 형성과정을 차례로 거치면서 선행되는 것이 관찰되었다. 이러한 사실은 세포막 구조의 변화와 이원형의 소실에 의하여 뒷받침이 될 수 있으으로 이러한 변화들을 융합과정에서의 각 단계로 해석하는 것이 가능하리라 사료된다.

• Mycobiology
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• 제29권1호
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• pp.15-18
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• 2001

Protoplast fusion is a useful technique for establishing fungal hybrids to overcome the natural barriers. The ultrastructure of protoplast and its fusion process were observed using a scanning electron microscopy(SEM) and a transmission electron microscopy(TEM). The protoplasts were variable in size from $0.5{\sim}15{\mu}m$ in diameter, and the mean diameter was about $3{\sim}5{\mu}m$. It was impossible to discriminate protoplasts of Lentinula edodes from protoplasts of Coriolus versicolor by size and surface structure. Big aggregates of the dehydrated protoplasts were observed, after polyethylene glycol 4000 treatment. Nucleus, mitochondria, lipid granules and various vesicles having granules were scattered in the cytoplasm. The vesicles were heterogeneous in size and vary from one protoplast to another. The fused membrane layer of the two protoplasts was observed. Time protoplast membrane contact and reorganization of membrane components were essential condition for protoplast fusion. Transmission electron micrograph showed fused protoplasts and flattening of the cells in the area of the membrane contact. We hope that our electron microscopic observations provide some insights into the understanding of the fusion process of protoplast in fungi.

• 미생물학회지
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• 제32권4호
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• pp.280-284
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• 1994

공생 아메바에서 리소솜과 공생낭 간에 융합이 저해되는 이유로서는 먼저 이들 공생낭의 막에 어떤 특별한 인자가 존재하여 융합을 저해하거나 또는 융합 과정에 필수적인 어떤 요소가 이들 공생막에는 부족하여 융합이 일어나지 않는다고 유추해 볼 수 있다. 단일 클론 항체를 추적물질로 사용하여 이들 인자나 구성요소를 알아내는 과정에서, lipopolysaccharides가 공생 박테리아에 의하여 생산되어 공생낭의 막에 삽입된다는 것을 확인하였으며 이들이 공생막상에서도 세포질 방향으로 노출되어 있다는 것을 알아내었다. 따라서 이들 lipopolysaccharides가 리소솜과 공생낭간의 융합 저해에 간여하는 가를 알아보기 위하여 이들에 대한 단일 크론 항체를 공생 아메자의 세포질에 미세주사하여 보았다. 주사된 아메바에서는 공생낭과 리소솜간의 융합이 일어나는 것으로 미루어 보아, 아마도 lipopolysaccharides는 융합저해 요소 중의 하나로 사료되어 진다.

• 한국동물학회지
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• 제38권4호
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• pp.483-489
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• 1995

많은 세포와 조직에서 분화 조절물질로 알려져 있는 비타민 A의 대사산물인 rectinoic acid(RA)가 배양 계배 근원세포의 성장과 분화에 미치는 영향을 조사하였다. RA가 처리된 세포는 그 농도가 증가함에 따라 융합 억제 효과도 증가함을 보였고, 배양액으로부터 RA를 제거해 주면 근원세포의 융합이 재개되는 것으로 보아, RA는 근원세포의 융합을 가역적으로 억제함을 알 수 있었다. 그러나 이미 최종 분화 단계에 들어선 세포에서는 융합 억제 효과를 보이지 않아, RA의 효과는 분화 단계에 따라 특이적으로 작용함을 보여준다. 융합 억제 효과에도 불구하고, RA는 근원세포의 융합 전단계인 방추형으로 신장하는 것과 그들이 가상의 축을 따라 배열하는 데는 영향을 주지 않았다. 또한 세포의 증식과 creatine kimase, tropomyosin과 같은 근특이 단백질의 축적에도 큰 영향을 주지 않았다. 한편, 근원세포는 융합하기 전에 필수적으로 fibronectin의 감소를 억제함을 알 수 있었다. 이상의 결과로 RA는 배양 근원세포의 융합만을 특이하게 억제하는 물질이며, 그 작용은 fibronectin의 감소를 방해하는 것과 밀접한 관계가 있는 것으로 사료된다.

• Toxicological Research
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• 제12권2호
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• pp.155-161
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• 1996

The enzymatic properties for NADPH-P450 reductase domain of a fusion protein between human cytochrome P450 1A1 and rat NADPH-P450 reductase expressed in Escherichia coli were investigated. The fusion plasmid pCW/1A1OR-expressed E. coli membrane showed high NADPH-cytochrome c reductase activity ($830.1\pm 85.8 nmol\cdot min^{-1}\cdot mg protein^{-1}$), while pCW control vector and P 450 1A1 expression vector pCW/1A1 showed relatively quite low activity ($4.35\pm 0.49, 3.27\pm 0.50 nmol\cdot min^{-1}\cdot mg protein^{-1}$, respectively). The kinetic curves for NADPH-cytochrome c reductase followed typical Michaelis-Menten kinetics. The $K_{max}$ and $V_{max}$ for NADPH-dependent reductase activity were $8.24\pm 2.61\mu $and $817.9\pm 60.8 nmol\cdot min^{-1}\cdot mg protein^{-1}$, respectively, whereas those for cytochrome c-dependent reductase activity were $19.97\pm 2.86\mu M$ and $1303.5\pm 67.1 nmol\cdot min^{-1}\cdot mg protein^{-1}$. The reductase activities were also compared with those of rat, porcine and human liver microsomes. The activity of pCW/ 1A1OR-expressed E. coli membrane was 15.2-fold higher than that of rat liver microsome. Treatment with benzo(a)pyrene, 7-ethoxyresorufin and $\alpha$-naphthofiavone which are known as specific substrates or inhibitor for human P450 1A1 increased NADPH-cytochrome c reductase activity of fusion protein in E. coli membrane dose-dependently. These results demonstrate that the membrane topology of fused enzyme may be important for activity of its NADPH-P450 reductase domain.

• 한국수소및신에너지학회논문집
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• 제32권4호
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• pp.219-227
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• 2021

Hydrogen isotopes, which are used as raw materials in fusion reaction, participate in the reaction only in small amount, and most of them are released together with impurities. In order to recover and reuse only hydrogen isotopes from this exhaust gas, a recovery process is required, and most of the hydrogen isotopes can be recovered using a Pd Membrane. In this study, the recovery rate of hydrogen isotopes was measured through the first and second stage Pd membrane experiments. In the case of the experiment using a single stage Pd membrane, about 99.2%, and in the case of the first stage and second stage Pd membrane connection experiments, a recovery rate of 99.9% or more was obtained. Therefore, the recovery rate of Pd membrane process applied to hydrogen can be applied to hydrogen isotopes. In addition, the simulation model was established using aspen custom modeler, a commercial software, and the validity of the simulation was checked by applying the references and experimental data. The simulation results based on the experimental data showed a difference of 2% or less.