Structural domains involved in the regulation of transmitter release by synapsins.
Synapsins are a family of neuron-specific phosphoproteins that regulate neurotransmitter release by associating with synaptic vesicles. Synapsins consist of a series of conserved and variable structural domains of unknown function. We performed a systematic structure-function analysis of the various domains of synapsin by assessing the actions of synapsin fragments on neurotransmitter release, presynaptic ultrastructure, and the biochemical interactions of synapsin. Injecting a peptide derived from domain A into the squid giant presynaptic terminal inhibited neurotransmitter release in a phosphorylation-dependent manner. This peptide had no effect on vesicle pool size, synaptic depression, or transmitter release kinetics. In contrast, a peptide fragment from domain C reduced the number of synaptic vesicles in the periphery of the active zone and increased the rate and extent of synaptic depression. This peptide also slowed the kinetics of neurotransmitter release without affecting the number of docked vesicles. The domain C peptide, as well as another peptide from domain E that is known to have identical effects on vesicle pool size and release kinetics, both specifically interfered with the binding of synapsins to actin but not with the binding of synapsins to synaptic vesicles. This suggests that both peptides interfere with release by preventing interactions of synapsins with actin. Thus, interactions of domains C and E with the actin cytoskeleton may allow synapsins to perform two roles in regulating release, whereas domain A has an actin-independent function that regulates transmitter release in a phosphorylation-sensitive manner.
Hilfiker
Sabine
S
Laboratory of Molecular and Cellular Neuroscience, The Rockefeller University, New York, New York 10021, USA. sabine.hilfiker@ipb.csic.es
Benfenati
Fabio
F
Doussau
Frédéric
F
Nairn
Angus C
AC
Czernik
Andrew J
AJ
Augustine
George J
GJ
Greengard
Paul
P
D000595:Amino Acid Sequence / Q000502:physiology; D000818:Animals; D002417:Cattle; D019706:Excitatory Postsynaptic Potentials / Q000502:physiology; D066298:In Vitro Techniques; D049834:Loligo; D008969:Molecular Sequence Data; D018377:Neurotransmitter Agents / Q000378:metabolism*; D010446:Peptide Fragments / Q000235:genetics / Q000494:pharmacology; D017434:Protein Structure, Tertiary / Q000187:drug effects / Q000502:physiology; D051381:Rats; D013233:Stellate Ganglion / Q000187:drug effects / Q000378:metabolism / Q000502:physiology; D016704:Synapsins / Q000737:chemistry* / Q000378:metabolism* / Q000502:physiology
D018377:Neurotransmitter Agents; D010446:Peptide Fragments; D016704:Synapsins