Biochemical and Pharmacolosical study of the Accumulation of Eycitatory amino Acids in to Synaptic Vesicles.

突触小泡中刺激性氨基酸积累的生化和药理学研究。

• 批准号: 05671821
• 负责人: MORIYAMA Yoshinori
• 金额: $ 1.28万
• 依托单位: Osaka University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (C)
• 财政年份: 1993
• 资助国家: 日本
• 起止时间: 1993 至 1994
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-05671821/
• 关键词: Glutamate; Synaptic vesicle; Transmitter; Vacuolar ATPase; Newron; Transporter; 輸送系; 液胞型ATPase

Synaptic vesicles are storage subcelluar organelles for neurotransmitters in presynaptic nerve terminals and possess at least four different kinds of transporters for neurotransmitters : One of them, a glutamate transporter is very important transporter, because glutamate is an exicitatory amino acid transmitter. So far little is known about the glutamate transporter. In this study I investigated several aspects of the transporter : (1) purification, (2) specific inhibitors, (3) biochemical study on vacuolar ATPase as a primary pump for glutamate transport. During the past two years, I succeeded at least in part in carrying out the above projects : I established the procedure for solubilization, reconstitution and partial purification of glutamate transporter. I found that dibezylglutamate as a potent competitive inhibitor for the transporter. I determinied that there are binding sites for omeprazole and quinacrine mustard in subunit A of chromaffin granules vacuolar ATPase. By cross-linking technique, I obtained several useful information on subunit/subunit interaction of vacuolar ATPases. These results are important for understanding entire feature of vesicular glutamate transporter.

突触囊泡是突触前神经末梢中神经递质的储存亚细胞器，至少具有四种不同的神经递质转运体：其中谷氨酸转运体是非常重要的转运体，因为谷氨酸是一种兴奋性氨基酸递质。到目前为止，人们对谷氨酸转运蛋白知之甚少。本研究从以下几个方面对谷氨酸转运蛋白进行了研究：（1）谷氨酸转运蛋白的纯化;（2）谷氨酸转运蛋白的特异性抑制剂;（3）谷氨酸转运蛋白的主要泵-液泡膜ATP酶的生化研究。在过去的两年中，我成功地至少部分地进行了上述项目：我建立了谷氨酸转运蛋白的溶解，重建和部分纯化的程序。我发现谷氨酸二苄酯是一种有效的转运蛋白竞争性抑制剂。在嗜铬颗粒空泡ATP酶A亚单位上，有奥美拉唑和奎纳克林芥子气的结合位点。通过交联技术，获得了液泡ATP酶亚基间相互作用的一些有用信息。这些结果对了解囊泡谷氨酸转运体的整体特性具有重要意义。

项目成果

Moriyama.Y.et.al: "Eridence for a common bindirg site for omeprczole & NeM in subunit A of chromatlin granwle V-ATPsce" Biochem.Bioplys Res.Commun.196. 699-706 (1993)

Moriyama.Y.et.al：“奥美唑的常见 Bindirg 位点的证据

Uyama, T., Moriyama, Y., Futai, M., and Michibata, H.: "Immunological detection of vacuolar-type proton ATPase in vanadocytes of the Ascidian Ascidia sydneinsis samea." J.Exp.Zool.270. 148-154 (1994)

Uyama, T.、Moriyama, Y.、Futai, M. 和 Michibata, H.：“海鞘 Ascidia sydneinsis Samea 的钒细胞中液泡型质子 ATP 酶的免疫学检测。”

Moriyama, Y., Patel, V., Ueda, I., and Futai, M.: "Evidence for a common binding site for omeprazole and N-ethylmaleimide in subunit A of chromaffin granule vacuolar-type ATPase." Biochem.Biophys.Res.commun.196. 699-706 (1993)

Moriyama, Y.、Patel, V.、Ueda, I. 和 Futai, M.：“嗜铬颗粒液泡型 ATP 酶亚基 A 中奥美拉唑和 N-乙基马来酰亚胺共同结合位点的证据。”

Uyama T.et.al: "Immunological detection of vacuolar-type H^+-ATPsce in vanalocytes of Ascidiom." J.Eyp.Zoolosy. 270. 148-154 (1994)

Uyama T.et.al：“Ascidiom 细胞中液泡型 H^ -ATPsce 的免疫学检测。”

Yoshinori Moriyama et.al: "Evidence for a common binding site for omeprazole and N-ethylmaleimide in subunit A of obromaffin granule vacuolar-type ATPase" Biochemical Biophysical Research Communications.196. 699-706 (1993)

Yoshinori Moriyama 等人：“奥美拉唑和 N-乙基马来酰亚胺在奥布罗芬颗粒液泡型 ATP 酶亚基 A 中存在共同结合位点的证据”《生化生物物理研究通讯》196。