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

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。

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.