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

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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.