The synaptic vesicle protein SV2A is the binding site for the antiepileptic drug levetiracetam.
Here, we show that the synaptic vesicle protein SV2A is the brain binding site of levetiracetam (LEV), a new antiepileptic drug with a unique activity profile in animal models of seizure and epilepsy. The LEV-binding site is enriched in synaptic vesicles, and photoaffinity labeling of purified synaptic vesicles confirms that it has an apparent molecular mass of approximately 90 kDa. Brain membranes and purified synaptic vesicles from mice lacking SV2A do not bind a tritiated LEV derivative, indicating that SV2A is necessary for LEV binding. LEV and related compounds bind to SV2A expressed in fibroblasts, indicating that SV2A is sufficient for LEV binding. No binding was observed to the related isoforms SV2B and SV2C. Furthermore, there is a high degree of correlation between binding affinities of a series of LEV derivatives to SV2A in fibroblasts and to the LEV-binding site in brain. Finally, there is a strong correlation between the affinity of a compound for SV2A and its ability to protect against seizures in an audiogenic mouse animal model of epilepsy. These experimental results suggest that SV2A is the binding site of LEV in the brain and that LEV acts by modulating the function of SV2A, supporting previous indications that LEV possesses a mechanism of action distinct from that of other antiepileptic drugs. Further, these results indicate that proteins involved in vesicle exocytosis, and SV2 in particular, are promising targets for the development of new CNS drug therapies.
Lynch
Berkley A
BA
Department of Molecular and Cellular Biology, UCB Research Inc., 840 Memorial Drive, Cambridge, MA 02139, USA. berkley.lynch@ucb-group.com
Lambeng
Nathalie
N
Nocka
Karl
K
Kensel-Hammes
Patricia
P
Bajjalieh
Sandra M
SM
Matagne
Alain
A
Fuks
Bruno
B
D000818:Animals; D000927:Anticonvulsants / Q000378:metabolism*; D001665:Binding Sites; D001921:Brain / Q000166:cytology / Q000378:metabolism; D005347:Fibroblasts; D017353:Gene Deletion; D006801:Humans; D020128:Inhibitory Concentration 50; D007425:Intracellular Membranes / Q000378:metabolism; D000077287:Levetiracetam; D008562:Membrane Glycoproteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D051379:Mice; D018345:Mice, Knockout; D008970:Molecular Weight; D009419:Nerve Tissue Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D019920:Photoaffinity Labels; D010889:Piracetam / Q000031:analogs & derivatives / Q000378:metabolism*; D011233:Precipitin Tests; D011485:Protein Binding; D051381:Rats; D012640:Seizures; D013572:Synaptic Vesicles / Q000378:metabolism
D000927:Anticonvulsants; D008562:Membrane Glycoproteins; D009419:Nerve Tissue Proteins; D019920:Photoaffinity Labels; C585899:SV2C protein, human; C489614:Sv2a protein, mouse; C489618:Sv2a protein, rat; C076414:SV2A protein, human; D000077287:Levetiracetam; D010889:Piracetam