Calsyntenin-3 molecular architecture and interaction with neurexin 1a.

Authors:
Lu Z, Wang Y, Chen F, Tong H, Reddy MV, Luo L, Seshadrinathan S, Zhang L, Holthauzen LM, Craig AM, Ren G, Rudenko G.
In:
Source: J Biol Chem
Publication Date: (2014)
Issue: 289(50): 34530-42
Research Area:
Neurobiology
Gene Expression
Cells used in publication:
High Five
Species: Trichopulsia ni (Cabbage Looper)
Tissue Origin:
Culture Media:
Abstract
Calsyntenin 3 (Cstn3 or Clstn3), a recently identified synaptic organizer, promotes the development of synapses. Cstn3 localizes to the postsynaptic membrane and triggers presynaptic differentiation. Calsyntenin members play an evolutionarily conserved role in memory and learning. Cstn3 was recently shown in cell-based assays to interact with neurexin 1a (n1a), a synaptic organizer that is implicated in neuropsychiatric disease. Interaction would permit Cstn3 and n1a to form a trans-synaptic complex and promote synaptic differentiation. However, it is contentious whether Cstn3 binds n1a directly. To understand the structure and function of Cstn3, we determined its architecture by electron microscopy and delineated the interaction between Cstn3 and n1a biochemically and biophysically. We show that Cstn3 ectodomains form monomers as well as tetramers that are stabilized by disulfide bonds and Ca(2+), and both are probably flexible in solution. We show further that the extracellular domains of Cstn3 and n1a interact directly and that both Cstn3 monomers and tetramers bind n1a with nanomolar affinity. The interaction is promoted by Ca(2+) and requires minimally the LNS domain of Cstn3. Furthermore, Cstn3 uses a fundamentally different mechanism to bind n1a compared with other neurexin partners, such as the synaptic organizer neuroligin 2, because Cstn3 does not strictly require the sixth LNS domain of n1a. Our structural data suggest how Cstn3 as a synaptic organizer on the postsynaptic membrane, particularly in tetrameric form, may assemble radially symmetric trans-synaptic bridges with the presynaptic synaptic organizer n1a to recruit and spatially organize proteins into networks essential for synaptic function.