Structural analysis of Ca²⁺-binding pocket of synaptotagmin 5 C2A domain

Synaptotagmins constitute a family of multifunctional integral membrane proteins found predominantly on vesicles in neural and endocrine tissues. 17 isoforms of synaptotagmin family in mammals have been identified, 7 isoforms among them are known to be able to bind Ca²⁺ via their C2 domains. This study presents the crystal structure of the first C2 domain (C2A domain) of synaptotagmin 5 complexed with Ca²⁺ at 1.90Å resolution. Comparison of the Ca²⁺-binding pocket of synaptotagmin 5 C2A domain with other synaptotagmin C2 domains demonstrated that a serine residue locating at Ca²⁺-binding loop probably responsible to the conformational variation of Ca²⁺-binding pocket, and thus impacts the Ca²⁺-binding mechanism of C2 domain, which is verified by structural analysis of the serine mutant and Ca²⁺-binding assays via isothermal titration calorimetry. Alteration of Ca²⁺-binding mechanism might be correlated with different Ca²⁺ response rates of synaptotagmins, which is the basis of the functions of synaptotagmins in regulating various types of Ca²⁺-triggered vesicle-membrane fusion processes.