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完整後設資料紀錄
DC 欄位 | 值 | 語言 |
---|---|---|
dc.contributor.advisor | 潘建源 | |
dc.contributor.author | Ying-Shiuan Wang | en |
dc.contributor.author | 王英亘 | zh_TW |
dc.date.accessioned | 2021-06-15T05:43:17Z | - |
dc.date.available | 2013-08-20 | |
dc.date.copyright | 2010-08-20 | |
dc.date.issued | 2010 | |
dc.date.submitted | 2010-08-20 | |
dc.identifier.citation | 1. Aravind P, Chandra K, Reddy PP, Jeromin A, Chary KV, Sharma Y. (2008) Regulatory and structural EF-hand motifs of neuronal calcium sensor-1: Mg 2+ modulates Ca2+ binding, Ca2+-induced conformational changes, and equilibrium unfolding transitions. J Mol Biol. 376(4), 1100-1115
2. Dason JS, Romero-Pozuelo J, Marin L, Iyengar BG, Klose MK, Ferrus A, Atwood HL. (2009) Frequenin/NCS-1 and the Ca2+-channel alpha1-subunit co-regulate synaptic transmission and nerve-terminal growth. J Cell Sci 122, 4109-4121 3. Gall WE, Higginbotham MA, Chen C, Ingram MF, Cyr DM, Graham TR. (2000) The Auxilin-like phosphoprotein Swa2p is required for clathrin function in yeast. Curr Biol 10(21), 1349-1358 4. Greener T, Zhao X, Nojima H, Eisenberg E, Greene LE. (2000) Role of cyclin G-associated kinase in uncoating clathrin-coated vesicles from non-neuronal cells. J Biol Chem. 275(2), 1365-1370 5. Greener T, Grant B, Zhang Y, Wu X, Greene LE, Hirsh D, Eisenberg E. (2001) Caenorhabditis elegans Auxilin: a J-domain protein essential for clathrin-mediated endocytosis in vivo. Nat Cell Biol 3(2), 215-219 6. Kabbani N, Negyessy L, Lin R, Goldman-Rakic P, Levenson R. (2002) Interaction with neuronal calcium sensor NCS-1 mediates desensitization of the D2 dopamine receptor. J Neurosci. 22(19), 8476-8486 7. Kandachar V, Bai T, Chang HC. (2008) The clathrin-binding motif and the J-domain of Drosophila Auxilin are essential for facilitating Notch ligand endocytosis. BMC Dev Biol. 8:50 8. Lee DW, Zhao X, Eisenberg E, Greene LE. (2005) Depletion of GAK/Auxilin 2 inhibits receptor-mediated endocytosis and recruitment of both clathrin and clathrin adaptors. J Cell Sci 118, 4311-4321 9. Lemmon SK. (2001) Clathrin uncoating: Auxilin comes to life. Curr Biol 11(2), 49-52 10. Ma Y, Greener T, Pacold ME, Kaushal S, Greene LE, Eisenberg E. (2002) Identification of domain required for catalytic activity of Auxilin in supporting clathrin uncoating by Hsc70. J Biol Chem. 277(51), 49267-49274 11. Massol RH, Boll W, Griffin AM, and Kirchhausen T (2006) A burst of Auxilin recruitment determines the onset of clathrin-coated vesicle uncoating. Proc Natl Acad Sci U S A 103(27), 10265-10270 12. McFerran BW, Graham ME, Burgoyne RD. (1998) Neuronal Ca2+ sensor 1, the mammalian homologue of frequenin, is expressed in chromaffin and PC12 cells and regulates neurosecretion from dense-core granules. J Biol Chem 273(35), 22768-22772 13. Moncrief ND, Kretsinger RHC, Goodman M. (1990) Evolution of EF-hand calcium-modulated proteins. J Mol Evol. 30, 522-562 14. Morgan JR, Prasad K, Jin S, Augustine GJ, Lafer EM. (2001) Uncoating of clathrin-coated vesicles in presynaptic terminals: roles for Hsc70 and Auxilin. Neuron. 32(2), 289-300 15. Negyessy L., Goldman-Rakic PS. (2005) Subcellular localization of the dopamine D2 receptor and coexistence with the calcium-binding protein neuronal calcium sensor-1 in the primate prefrontal cortex. J Comp Neurol. 488(4), 464-475 16. Pan CY, Jeromin A, Lundstrom K, Yoo SH, Roder J, Fox AP. (2002) Alterations in Exocytosis Induced by Neuronal Ca2+ Sensor-1 in Bovine Chromaffin Cells. J Neurosci 22(7), 2427-2433 17. Pishvaee B, Costaguta G, Yeung BG, Ryazantsev S, Greener T, Greene LE, Eisenberg E, McCaffery JM, Payne GS. (2000) A yeast DNA J protein required for uncoating of clathrin-coated vesicles in vivo. Nat Cell Biol. 2(12), 958-963 18. Pongs O, Lindemeier J, Zhu XR, Theil T, Engelkamp D, Krah-Jentgens I, Lambrecht HG, Noch KW, Schwemer J, Rivosecchi R, Mallart A, Galceran J, Canal J, Barbas JA, and Ferrus A (1993) Frequenin-A Novel CalciumBinding Protein That Modulates Synaptic Efficacy in the Drosophila Nervous System. Neuron 11(1), 15-28 19. Prasad K, Barouch W, Greene L, Eisenberg E. (1993) A protein cofoactor is required for uncoating of clathrin baskets by uncoating ATPase. J Biol Chem 268(32), 23758-23761 20. Scheele U, Alves J, Frank R, Duwel M, Kalthoff C, Ungewickell E. (2003) Molecular and functional characterization of clathrin- and AP-2-binding determinants within a disordered domain of Auxilin. J Biol Chem 278(28), 25357-25368 21. Sudhof TC. (2004) The Synaptic Vesicle Cycle. Annu Rev Neurosci 27, 509-547 22. Tsai J, and Douglas MG. (1996) A conserved HPD sequence of the J-domain is necessary for YDJ1 stimulation of Hsp70 ATPase activity at a site distinct from substrate binding. J. Biol Chem 271(16), 9347-9354 23. Tsujimoto T, Jeromin A, Saitoh N, Roder JC, Takahashi T. (2002) Neuronal calcium sensor 1 and activity-dependent facilitation of P/Q-type calcium currents at presynaptic nerve terminals. Science 295(5563), 2276-2279 24. Weiss JL, Archer DA, Burgoyne RD. (2000) Neuronal Ca2+ sensor-1/frequenin functions in an autocrine pathway regulating Ca2+ channels in bovine adrenal chromaffin cells. J Biol Chem 275(51), 40082-40087 | |
dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/46918 | - |
dc.description.abstract | 神經傳遞物質以胞吐釋放後,同時也靠著胞吞作用,來以維持神經細胞細胞膜表面積的恆定。Neuron calcium sensor 1 (NCS-1)是EF-hand鈣離子結合蛋白家族的一員,其會與鈣離子結合並調控神經傳遞物質的釋放。此外,本實驗室之前的研究以Yeast-two-hybrid screening發現NCS-1會與Auxilin-1的C端(胺基酸序列為384至910)結合。Auxilin-1分布在神經細胞突觸上,在網格蛋白調控的胞吞機轉中,幫助網格蛋白從胞吞作用進來的小泡上離開。為更了解兩者扮演的角色,將NCS-1以及Auxilin-1基因在腎上腺嗜鉻細胞中大量表現並給予刺激,觀察鈣離子流入量及細胞膜電容變化。結果顯示,失去與鈣離子結合能力之NCS-1E120Q及無法與細胞膜鍵結的NCS-1G2A在連續刺激下,在鈣離子流入量有降低的趨勢,而NCS-1E120Q則使胞吐量顯著下降。而表現Auxilin-1的細胞,並不影響鈣離子流入量及膜電容變化。而失去與Hsc70結合能力的Auxilin-1H874Q,則使鈣離子流入量及胞吐作用上顯著下降。在胞吞能力上,NCS-1與Auxilin-1都有著較慢的速率。這些結果顯示NCS-1與鈣離子的結合,可幫助小泡來到Readily releasable pool (RRP),以準備釋放,並幫助細胞在刺激結束後,進行胞吞以維持細胞膜的恆定。Auxilin-1會使細胞RRP的量變小,但不影響總釋放量。而NCS-1與Auxilin-1在牛腎上腺嗜鉻細胞中,有共同的表現位置,與之前研究不同的地方在於NCS-1會參與在胞吞作用上,Auxilin-1會參與到胞吐作用上,因此NCS-1與Auxilin-1兩者之間可能有交互作用,共同調控胞吞胞吐作用。 | zh_TW |
dc.description.abstract | During synaptic cycle in neurotransmitter release, the surface area of cell membrane maintains homeostasis by exo/endocytosis. Neuron calcium sensor 1 (NCS-1), a member of EF-hand calcium-binding protein family, is shown to regulate neurotransmitter release. Our previous results have shown that the half part of C terminus (a.a. 444-970) in rat Auxilin-1 binds to NCS-1 by yeast-two-hybrid screening using NCS-1 as the bait. Auxilin-1 is expressed in synapse, and facilitates clathrin uncoating from clathrin-coated vesicles. To characterize the roles of NCS-1 and Auxilin-1 in exo/endocytosis, the effects of NCS-1 and Auxilin-1 on evoked exo/endocytosis were monitored by the calcium influx and capacitance measurements in bovine chromaffin cells. In confocal images, overexpressed NCS-1 and Auxilin-1 are colocalized in chromaffin cells. Both mutants NCS-1E120Q with the abolished calcium binding ability and NCS-1G2A with the abolished membrane anchoring ability decreased the calcium influx during repetitive stimuli. Furthermore, NCS-1E120Q significantly reduced exocytosis. In contrast, overexpression of human Auxilin-1 had no significant effect on calcium influx and exocytosis; however, Auxilin-1H874Q mutant which loses its ability to interact with Hsc70 significantly decreased the calcium influx and exocytosis. For endocytosis measured by capacitance, both NCS-1 and Auxilin-1 had a slower rate of membrane retrival. These results indicate that calcium binding to NCS-1 helps vesicles recycling to the plasma membrane, and Auxilin-1 takes part in the vesicle recycling pathway, but Auxilin-1H874Q decreases the endocytosis rate. In conclusion, NCS-1 may interact with Auxilin-1 on the half part of C terminus to regulate exo/endocytosis. | en |
dc.description.provenance | Made available in DSpace on 2021-06-15T05:43:17Z (GMT). No. of bitstreams: 1 ntu-99-R97B41016-1.pdf: 8153414 bytes, checksum: 0c7c6c91269b065bafb34f7a444ec693 (MD5) Previous issue date: 2010 | en |
dc.description.tableofcontents | 前言 I
突觸小泡循環機制 1 Neuron Calcium Sensor-1 (NCS-1) 1 網格蛋白與胞吞作用 3 Auxilin 4 研究目標 6 材料與方法 7 人類Auxilin-1基因片段製備 7 大鼠NCS-1基因片段製備 7 聚合酶連鎖反應(Polymerasae Chain Reaction, PCR) 8 TA cloning 8 連接作用(Ligation) 8 轉型作用(Transformation) 9 質體製備 9 點突變 9 初級牛腎上腺嗜鉻細胞培養 10 轉移感染(Transfection) 10 電生理紀錄-細胞膜電容測量 11 細胞免疫螢光染色法 12 資料分析 12 結果 14 NCS-1與Auxilin-1在牛嗜鉻細胞的分佈 14 NCS-1與Auxilin-1在刺激前細胞大小並無顯著差異 14 NCS-1突變株抑制鈣離子進入細胞內的能力 15 Auxilin-1的突變株降低鈣離子電流 16 NCS-1對於胞吐作用的影響 16 Auxilin-1對於胞吐作用的影響 17 NCS-1與Auxilin-1鈣離子依賴性 18 NCS-1與Auxilin-1對於胞吞作用的影響 19 NCS-1與Auxilin-1在細胞內的交互作用 19 討論 20 NCS-1與Auxilin-1的分佈 20 細胞大小之間的差異 21 NCS-1E120Q明顯的降低鈣離子流入量 21 NCS-1E120Q明顯抑制胞吐作用 22 Auxilin-1H874Q顯著降低了鈣離子流量和胞吐作用 23 NCS-1與Auxilin-1在鈣離子依賴性的影響 24 NCS-1與Auxilin-1對胞吞作用的影響 24 NCS-1與Auxilin-1有交互作用 25 結論 26 參考文獻 27 表 31 圖解 33 圖 37 | |
dc.language.iso | zh-TW | |
dc.title | NCS-1與Auxilin-1對胞吞/胞吐作用之影響 | zh_TW |
dc.title | Effects of NCS-1 and Auxilin-1 on Exo/endocytosis | en |
dc.type | Thesis | |
dc.date.schoolyear | 98-2 | |
dc.description.degree | 碩士 | |
dc.contributor.oralexamcommittee | 戴晶瑩,王致恬,林崇智 | |
dc.subject.keyword | NCS-1,Auxilin-1,胞吞胞吐作用,鈣離子,腎上腺嗜鉻細胞, | zh_TW |
dc.subject.keyword | NCS-1,Auxilin-1,exo/endocytosis,calcium,chromaffin cell, | en |
dc.relation.page | 48 | |
dc.rights.note | 有償授權 | |
dc.date.accepted | 2010-08-20 | |
dc.contributor.author-college | 生命科學院 | zh_TW |
dc.contributor.author-dept | 動物學研究所 | zh_TW |
顯示於系所單位: | 動物學研究所 |
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