• 제목/요약/키워드: F-actin

검색결과 95건 처리시간 0.027초

항-액틴-금 입자 표지에 의한 개불(Urechis unicinctus) 정자 및 정세포 핵 Actin의 분포 (Localization of Anti-Actin-Gold Particles (10 nm) Labeled to Nuclear Actin of Urechis Sperm and Spermatids)

  • 신길상;김호진;김완종
    • Applied Microscopy
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    • 제30권4호
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    • pp.403-412
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    • 2000
  • 1. 아메바 항-액틴의 Ag-Ab 반응과 이를 항-생쥐 IgG-금 입자로 표지한 결과는 주로 정세 포 및 정자의 핵질에 특이적으로 표지되었고 첨체에서는 그 반응을 볼 수 없었다. 2. 표지된 항-액탠 금 입자로 볼 때 정자 핵질의 G-액틴 또는 G-actin oligomer는 정세포의 F-액틴에서 유래되는 것으로 사료된다. 3. 첨체의 액틴은 주로 F-액틴으로 첨체돌기 형성에 참여하지 않는 위상인 것으로 관찰된다. 4. 미세구조의 변화, 정세포 체적의 감소, 정세포 및 정자 핵에 표지되는 금 입자의 증가와 이들 현상이 나타나는 동시성으로 볼 때 정세포 핵의 형태변화의 내용은 응축이고 이는 F-actin의 탈중합 반응에 의한 G-액틴의 생성이 원인일 것으로 사료된다.

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수축중인 근육막에서 actin과 myosin 결합의 구조변화에 관한 연구 (A Study On Structure Change of Binding Actin and Moysin On The Contracting Muscle Membrane)

  • 김덕술;박근호
    • 한국응용과학기술학회지
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    • 제25권3호
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    • pp.380-387
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    • 2008
  • The effects of the applied stretch and MgADP binding on the structure of the actin and myosin cross-bridges in rabbit fibers in the rigor state have been investigatedwith improved resolution by x-ray diffraction using synchrotron radiation. To clarify the structure of the ATP hydrolysis intermediates formed by actin and myosin cross-bridges,the effects of various phosphate analogs in the of MgADP on the structure of the thin and thick filaments in glycerinated rabbit muscle fibers in the rigor state investigated by x-ray diffraction with a short exposure time using synchrotron radiation. These results strongly suggest that when MgADP and phosphate analogs such as metallofluorides(BeF3 and AlF4)and vanadate(VO4(Vi)) were added the rigor fibers in the presence of the ATP-depletion backup system, the intensities of the actin-based layer lines were markedly weakened. We found that the intensity of the 14.5 nm-based meridional reflections increase by 20-50% when phosphate analogs such as metallofluorides(BeF3 and AlF4) and vanadate(VO4(Vi)) was added to the rigor muscle.

Agrobactrium tumefaciens KU12로부터 분리한 pli12의 Replication Origin과 벼의 Actin 유전자 프로모터를 이용한 벼의 Binary Vector 제조 (Construction of Binary Vectors for the Rice Transformation Using a Rice Actin Promoter and Replication Origin of pTi12 Isolated from Agrobacterium tumefaciens KU12)

  • Sim, Woong-Seop
    • Journal of Plant Biology
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    • 제38권4호
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    • pp.365-371
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    • 1995
  • Binary vectors, pBI-ActR1, pBI-ActF1 and pBSH-ActR1, were constructed using pGA642, the replication origin of pTi12 and the rice actin promoter. The sizes of pBI-ActR1, pBI-ActF1 and pBSH-ActR1 were 12.9 kb, 13.2 kb and 11.95 kb, respectively. These vectors containing a rice actin promoter followed by a GUS structural gene could induce stronly the expression of GUS gene in transformed rice cells. Rice explants from 3-4 day old seedlings after germinatin were cocultured with A. tumefaceins harboring pBI-ActR1, pBI-ActF1 or pBSH-ActR1, and then GUS expression in the explants was assayed. Transformation of rice explants by these binary vectors was tissue-specific, such that the meristematic regions of shoot apex, root and hypocotyl were transformed by these binary vectors.

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F-actin cytoskeleton이 Jurkat T 림파구의 microvilli 형성에 미치는 영향 (Involvement of F-Actin Cytoskeleton for Microvilli Formation of Jurkat T Lymphocyte)

  • 이재설;김해영;손기애;김지은;문경미;김광현;최은봉;이종환
    • 생명과학회지
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    • 제21권10호
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    • pp.1401-1406
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    • 2011
  • 면역세포는 외부 병원체 감염, 자연적 순환에 대하여 형태변화를 수반한다. T세포는 염증, 면역 감시, 이동, 그리고 혈관통과를 위해 uropod, filopodia, lamellipodia, 및 microvilli를 생산한다. 짧고 손가락 처럼 생긴 microvilli는 순환하고 있는 포유동물 면역세포 표면을 덮고 있다. 단핵세포와 호중구의 세포표면은 많이 다른데 membrane ruffle을 함유하고 있다. 본 연구는, T세포의 microvilli에 대하여 actin cytoskeleton과의 연관성에 대하여 탐구하였다. Actin 파괴자인 cytochalasin D 처리 후 SEM관찰을 통해서, Jurkat T세포의 microvilli를 보면 빠르게 사라지는 것을 알 수 있었다. 이와는 대조적으로 RhoA의 activator인 PMA는 LIMK와 cofilin 신호 전달을 통해서 microvilli 두께가 확장되는 것을 관찰 하였다. 또한, cytochalasin D 처리는 EL4 T세포의 극성을 사라지게 하는 것으로 보아 F-actin은 T세포의 극성 유지에도 영향을 미친다. 이상의 결과는 Actin cytoskeleton은 T세포에서 microvilli와 극성 유지에 관여하고 있는 것을 제시한다.

아날로그에 효과에 의한 토끼 근육 막 수축의 중간단계 연구 (A Study on the Middle Step of Rabbit Skeletal Muscle Membrane Contraction by Analog Effects)

  • 김덕술
    • 한국응용과학기술학회지
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    • 제24권1호
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    • pp.61-66
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    • 2007
  • X-ray diffraction studies have been made to investigate the effects of binding of ADP, ADP+Vi, ADP+AIF4, $ADP+BeF_3$ on the structure of glycerinated rabbit skeletal muscle in the rigor state. Although these phosphate analogs are known to bind actively cycling myosin heads, it is not clear whether they can bind to the attached heads in the rigor muscle. We have found that these analogs can bind to the myosin heads attached to actin filaments in the rigor state. The present results indicate that (1) bound myosin heads altered their conformation in the proximal end toward the plane perpendicular to the fiber axis when MgADP bound to them, and (2) myosin heads were dissociated substantially (up to 50%) from actin filaments but still remained in the vicinity of actin filaments when MgADP and metallofluorides (AIF4 and BeF3) or vanadate bound to them. We detected new conformations of myosin heads attached to actin filaments when they had MgADP or ADP.Pi analogs. We report here these findings on the effects of MgADP and MgADP+phosphate analogs to the rigor crossbridges.

Repression of the F-box protein Skp2 is essential for actin damage-induced tetraploid G1 arrest

  • Jo, Yongsam;Shin, Deug Y.
    • BMB Reports
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    • 제50권7호
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    • pp.379-383
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    • 2017
  • We previously reported that p53 plays a role as a key regulator in the tetraploid G1 checkpoint, which is activated by actin damage-induced cytokinesis blockade and then prevents uncoupled DNA replication and nuclear division without cytokinesis. In this study, we investigated a role of Skp2, which targets CDK2 inhibitor p27/Kip1, in actin damage-induced tetraploid G1 arrest. Expression of Skp2 was reduced, but p27/Kip1 was increased, after actin damage-induced cytokinesis blockade. The role of Skp2 repression in tetraploid G1 arrest was investigated by analyzing the effects of ectopic expression of Skp2. After actin damage, ectopic expression of Skp2 resulted in DNA synthesis and accumulation of multinucleated cells, and ultimately, induction of apoptosis. These results suggest that Skp2 repression is important for sustaining tetraploid G1 arrest after cytokinesis blockade and is required to prevent uncoupled DNA replication and nuclear division without cytokinesis.

Novel Macrolide Actin-inhibitors Isolated from Sea Sponges

  • Karaki, Hideaki;Ozaki, Hiroshi
    • Toxicological Research
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    • 제17권
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    • pp.105-108
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    • 2001
  • Several marine toxins with macrolide structure have been found to act on actin. One of these toxins is mycalolide B isolated from the genus Mycale. This compound belongs to macrolide antibiotics and consists of tris-oxazole with strong cytotoxic activity ($IC_{50}$: 10-50 nM for growth of L1210 murine leukemia cells). This compound was found to be an actin-depolymerizing agent with the mode of action distinct from that of the known actin inhibitor, cytochalasin D. Tolytoxin, a macrolide isolated from cyano-bacteria with similar chemical structure to mycalolide B, seems to have similar effect. Another macrolide compound, aplyronine A, showed the effects similar to those of mycalolide B. Although bistheonellide A, a dimeric macrolide, did not show a severing effect, it de polymerized F-actin and sequestered G-actin by forming 1 : 2 complex with G-actins. Swinholide A has a structure and effects similar to those of bistheonel-lide A. In conclusion, mycalolide B, tolytoxin, aplyronine A, bistheonellide A and swinholide A are the members of "actin de polymerizing macrolide" the mechanism of which is different from that of cytochalasin D.halasin D.

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Swiprosin-1 Regulates Cytokine Expression of Human Mast Cell Line HMC-1 through Actin Remodeling

  • Ramesh, T.P.;Kim, Young-Dae;Kwon, Min-Sung;Jun, Chang-Duk;Kim, Sang-Wook
    • IMMUNE NETWORK
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    • 제9권6호
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    • pp.274-284
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    • 2009
  • Background: Swiprosin-1 was identified in human CD8+ lymphocytes, mature B cells and non-lymphonoid tissue. We have recently reported that swiprosin-1 is expressed in mast cells and up-regulated in both in vitro and in vivo. Methods: The expression of cytokines and swiprosin-1 were determined by by real time PCR and conventional PCR. Pharmacological inhibitors were treated to investigate potential mechanism of swiprosin-1 in mast cell activation. Actin content was evaluated by confocal microscopy and flow cytometry. Results: The swiprosin-1 augmented PMA/A23187-induced expression of cytokines and release of histamine. However, knock-down of swiprosin-1 showed only a modest effect on PMA/A23187-induced cytokine expression, suggesting that swiprosin-1 has gain-of-function characteristics. Swiprosin-1 was found in microvilli-like membrane protrusions and highly co-localized with F-actin. Importantly, either disruption of actin by cytochalasin B or inhibition of PI3 kinase, an enzyme involved in actin remodeling, by wortmannin blocked cytokine expression only in swiprosin-1-overexpressing cells. Conclusion: These results suggest that swiprosin-1 modulates mast cell activation potentially through actin regulation.

Quercetin Reduces Chemotactic Activity of Porcine Peripheral Blood Polymorphonuclear Cells

  • Hwa, Gyeong-Rok;Ahn, Changhwan;Kim, Hakhyun;Kang, Byeong-Teck;Jeung, Eui-Bae;Yang, Mhan-Pyo
    • 한국임상수의학회지
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    • 제39권2호
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    • pp.51-58
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    • 2022
  • Quercetin, a flavonoid found in fruits and vegetables, exhibits a strong anti-inflammatory activity. The objective of this study was to examine the effect of quercetin on chemotactic activity of peripheral blood polymorphonuclear cells (PMNs) to culture supernatant from peripheral blood mononuclear cells (PBMCs) stimulated with lipopolysaccharide (LPS). In addition, we determined whether this effect is related to interleukin (IL)-8 and changes in cytoskeleton. The chemotactic activity of PMNs was evaluated by a modified Boyden chamber assay. Total cellular filamentous (F)-actin levels were measured by method of fluorescence microscopy. The levels of IL-8 mRNA and protein were measured by real time polymerase reaction method and enzyme-linked immunosorbent assay, respectively. Quercetin (0-50 µM) itself has no chemoattractant effect for PMNs. The culture supernatant from PBMCs (2 × 106 cells/mL) treated with LPS (1 ㎍/mL) showed remarkable increase in chemotaxis of PMNs. However, this effect was reduced dose-dependently by treatment with quercetin. In addition, PBMCs treated with LPS revealed enhanced levels in IL-8 protein and mRNA. Co-treatment of LPS with quercetin (50 µM) in PBMCs decreased IL-8 production and expression. Treatment of quercetin (0-50 µM) on PMNs to rpIL-8 (10 nM) decreased dose-dependently the chemotactic activity of PMNs. Treatment of quercetin on PMNs to IL-8 also reduced their total cellular F-actin level. These results suggested that quercetin attenuates chemotactic activity of PMNs, which is mediated by down-regulation of IL-8 production from LPS-stimulated PBMCs and inhibition of F-actin polymerization in PMNs.