Calcium Binding Promotes Conformational Flexibility of the Neuronal Ca2+ Sensor Synaptotagmin

Synaptotagmin 1 (Syt1) is a synaptic vesicle protein that serves as a calcium sensor of neuronal secretion. It is established that calcium binding to Syt1 triggers vesicle fusion and release of neuronal transmitters, however, the dynamics of this process is not fully understood. To investigate how Ca2+ binding affects Syt1 conformational dynamics, we performed prolonged molecular dynamics (MD) simulations of Ca2+-unbound and Ca2+-bound forms of Syt1. MD simulations were performed at a microsecond scale and combined with Monte Carlo sampling. We found that in the absence of Ca2+ Syt1 structure in the solution is represented by an ensemble of conformational states with tightly coupled domains. To investigate the effect of Ca2+ binding, we used two different strategies to generate a molecular model of a Ca2+-bound form of Syt1. First, we employed subsequent replacements of monovalent cations transiently captured within Syt1 Ca2+-binding pockets by Ca2+ ions. Second, we performed MD simulations of Syt1 at elevated Ca2+ levels. All the simulations produced Syt1 structures bound to four Ca2+ ions, two ions chelated at the binding pocket of each domain. MD simulations of the Ca2+-bound form of Syt1 revealed that Syt1 conformational flexibility drastically increased upon Ca2+ binding. In the presence of Ca2+, the separation between domains increased, and interdomain rotations became more frequent. These findings suggest that Ca2+ binding to Syt1 may induce major changes in the Syt1 conformational state, which in turn may initiate the fusion process.