ATP Binding and Turnover in Drosophila Synapsin: Cloning and Characterization of the Synapsin 1 C-Domain

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Authors

Yee, Michelle

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2010

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Thesis

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en_US

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Abstract

Synapsins are phosphoproteins in the brain located on synaptic vesicles and involved with regulating neurotransmitter release. The synapsins regulate the amount of synaptic vesicles available for release of neurotransmitters into the synapse through exocytosis and recycling mechanisms. Synapsin is evolutionarily conserved among species, especially in the large central C-domain. In mice lacking synapsin, neurons rapidly run out of synaptic vesicles, indicating that synapsin plays a key role in neuronal transmission. Synapsin mutations have been found in families with genetic forms of epilepsy. Although the function of synapsin is unknown, it has been shown that the C-domain binds ATP. Experimental evidence indicates that the residue at position 373 has a key role in ATP binding. However, in Drosophila, there is a serine substituted for glutamic acid at this position. Therefore, synapsin may not bind ATP. The goal of this project is to study whether or not the Drosophila synapsin C-domain binds ATP. Drosophila synapsin will be cloned and expressed as a GST fusion protein, purified, and analyzed with stop-flow techniques. The results of these experiments will establish whether or not ATP binding is essential for synapsin function.

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