Molecular architecture of a prototypical trans synaptic complex GluD2 Cerebelli...
Molecular architecture of a prototypical trans synaptic complex GluD2 Cerebellin1 Neurexin1
Chemical synapses in the central nervous system (CNS) employ a multitude of neuronal cell-surface receptors, adhesion proteins, secreted effector molecules, and proteoglycans. Within such vast network of interactions, synapse-span...
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Descripción del proyecto
Chemical synapses in the central nervous system (CNS) employ a multitude of neuronal cell-surface receptors, adhesion proteins, secreted effector molecules, and proteoglycans. Within such vast network of interactions, synapse-spanning protein complexes mediate cell-cell adhesion, align pre- and postsynaptic specializations and exert bidirectional signaling, inducing pre- and postsynaptic differentiation. This proposal focuses on the Glutamate receptor D2–Cerebellin1–Neurexin1β trans-synaptic complex, a key component of the excitatory parallel fiber - Purkinje cell (PF-PC) synapse in the cerebellum. This tripartite interaction is essential for bidirectional synaptogenesis, and its specific disruption leads to impairment of cognition and motor coordination. However, the structure of this complex and its implications for synapse organization and modulation of neurotransmission remain unknown.
The goal of this proposal is to elucidate the molecular architecture of the GluD2–Cerebellin1–Neurexin1β protein complex, and understand its functional implications. I will use X-ray crystallography to obtain high-resolution structural information on binary and ternary complexes between the soluble, extracellular GluD2, Cerebellin1 and Neurexin1β regions. A range of biophysical methods combined with site-directed mutagenesis will be applied to dissect complex formation with respect to affinity, kinetics, stoichiometry and contribution of functional modules. In parallel, using cryo-electron tomography I aim to visualize and reconstruct the higher-order architecture of GluD2–Cerebellin1, and ultimately the trans-synaptic triad, in model cellular membranes. This integrated approach should reveal general principles of supra-molecular organization and function at neuronal synapses, as structurally related molecules are broadly present within the CNS. Understanding synaptic functions in molecular terms will produce enduring paradigms in basic neuroscience and benefit human health.