AI Article Synopsis
- * Researchers used a pull-down assay to identify binding proteins, finding that NAP-22 interacts with glutamic acid decarboxylase (GAD) isoforms GAD65 and GAD67.
- * The study suggests that NAP-22 may help transport GAD enzymes to synaptic regions and keep them anchored to synaptic vesicles, without affecting their enzymatic activity.
Article Abstract
NAP-22 (also called BASP1 or CAP-23) is a neuron-enriched protein localized mainly in the synaptic vesicles and the synaptic plasma membrane. Biochemically, it is recovered in the lipid raft fraction. In order to understand the physiological function of the neuronal lipid raft, NAP-22 binding proteins were screened with a pull-down assay. Glutamic acid decarboxylase (GAD) was detected through LC-MS/MS, and Western blotting using a specific antibody confirmed the result. Two isoforms of GAD, GAD65 and GAD67, were expressed in bacteria as GST-fusion forms and the interaction with NAP-22 was confirmed in vitro. Partial co-localization of NAP-22 with GAD65 and GAD67 was also observed in cultured neurons. The binding showed no effect on the enzymatic activity of GAD65 and GAD67. These results hence suggest that NAP-22 could participate in the transport of GAD65 and GAD67 to the presynaptic termini and their retention on the synaptic vesicles as an anchoring protein.
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| http://dx.doi.org/10.1016/j.neulet.2013.01.030 | DOI Listing |
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