Molecular Mechanisms of Exocytotic Proces and Its Regulation

胞吐过程的分子机制及其调控

• 批准号: 12480232
• 负责人: KUMAKURA Konosuke
• 金额: $ 9.02万
• 依托单位: SOPHIA UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 2000
• 资助国家: 日本
• 起止时间: 2000 至 2002
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-12480232/
• 关键词: Chromaffin Cell; Exocytosis; Fusion kinetics; Vesicle movement; PKC; actin-myosin interaction; actin cytoskeleton; SNARE complex; カーボンファイバー; イノシトール-6-リン酸

The aim of this research project is to elucidate the molecular mechanism of exocytotic process and its regulation and we have obtained the following results.1)Roles of PKC in vesicle recruitment and priming.The facilitation of the priming by protein kinase C (PKC) is mediated through the complex between the activated PKC a or PKC 13, their receptor molecule RACK 1 and cortical F-actin. Vesicle recruitment was also regulated by PKC through the vesicle movement at the rate of 0.03-0.06 μm/s5 and the facilitation of vesicle recruitment was detected as an increase in the frequency of exocytotic events.2)The mechanism of priming and roles of actin cytoskeleton.The inhibition of actin-myosin interaction abolished the recruitment of chromaffin granules to the release sites, probably by reducing the population of rapidly moving granules that consist of about 25% of vesicles.3)Regulation of SNARE complex.Binding of calmodulin to C-teminus of VAMP-2, and binding between Syntaxin 1A and CaMkinase II regulate SNARE complex and, are essential for exocytosis.

本研究旨在阐明胞吐过程及其调控的分子机制，得到以下结果：1)蛋白激酶C(PKC)在囊泡募集和启动过程中的作用，蛋白激酶C(PKC)通过激活的PKC a或PKC 13与其受体RAC1和皮质F-肌动蛋白之间的复合体介导启动。蛋白激酶C还通过囊泡以0.03-0.06μm/S5的速率运动来调节囊泡的募集，并通过增加胞吐事件的频率来促进囊泡的募集。2)启动的机制和肌动蛋白细胞骨架的作用。抑制肌球蛋白的相互作用取消了嗜铬颗粒在释放部位的募集，可能是通过减少快速移动的颗粒的数量，这些颗粒约占囊泡总数的25%。3)SNARE复合体的调节。钙调蛋白与VAMP-2的C-teminus结合，以及合成酶1A和CaMK II之间的结合调节SNARE复合体，这对胞吐是必不可少的。

项目成果

伊藤 尚: "Increase in N-terminal hydrophobicity in mastoparan molecule enhanced the original hemolytic and catecholamine releasing activities from chromaffin cells."東京保健科学学会誌. 3(2). 109-117 (2000)

Hisashi Ito：“mastoparan 分子 N 末端疏水性的增加增强了嗜铬细胞的原始溶血和儿茶酚胺释放活性。” 东京健康科学学会杂志 3(2) (2000)。

Kenjiro Noyama, Shohei Maekawa: "Localization of cyclic nucleotide phosphodiesterase 2 in the brain-derived Triton-insoluble low-density fraction(raft)"Neurosci.Res.. 45. 141-148 (2003)

Kenjiro Noyama、Shohei Maekawa：“环核苷酸磷酸二酯酶 2 在脑源性 Triton 不溶性低密度组分（筏）中的定位”Neurosci.Res.. 45. 141-148 (2003)

Stephanie Quetglas, Cecile Iborra, Nobuyuki Sasakawa, Luc De Haro, Konosuke Kumakura, Kazuki Sato, Christian Leveque, Michael Seagar: "Calmodulin and lipid binding to synaptovrevin regulates calcium-dependent exocytosis"The EMBO Journal. 21. 1-10 (2002)

Stephanie Quetglas、Cecile Iborra、Nobuyuki Sasakawa、Luc De Haro、Konosuke Kumakura、Kazuki Sato、Christian Leveque、Michael Seagar：“钙调蛋白和脂质与 synaptovrevin 结合调节钙依赖性胞吐作用”EMBO 杂志。

Akihiro Ohyama, Kohei Hosaka, Yoshiaki Komiya, Kimio Akagawa, Emiko Yamauchi, Hisaaki Taniguchi, Nobuyuki Sasakawa, Konosuke Kumakura, Sumiko Mochida, Takashi Yamauchi, Michihiro Igarashi: "Regulation of exocytosis through Ca^<2+>/ATP-dependent binding of

Akihiro Ohyama、Kohei Hosaka、Yoshiaki Komiya、Kimio Akakawa、Emiko Yamauchi、Hisaaki Taniguchi、Nobuyuki Sasakawa、Konosuke Kumakura、Sumiko Mochida、Takashi Yamauchi、Michihiro Igarashi：“通过 Ca^<2>/ATP 依赖性结合调节胞吐作用

Ohira,K.: "Absence of Trk B and Insulin receptor β in the Triton insolube low-density fraction (raft) of rat brain."NeuroReport. 11. 1307-1311 (2000)

Ohira, K.：“大鼠大脑 Triton 不溶低密度部分（筏）中缺乏 Trk B 和胰岛素受体 β。”NeuroReport。11. 1307-1311 (2000)