Allosteric stabilization of calcium and phosphoinositide dual binding engages several synaptotagmins in fast exocytosis

• Published in: eLife 2022-08, Vol.11
• Main Authors: Kobbersmed, Janus RL, Berns, Manon MM, Ditlevsen, Susanne, Sørensen, Jakob B, Walter, Alexander M
• Format: Article
• Language: English
• Subjects: Affinity; Allosteric properties; allostericity; Binding sites; Calcium (intracellular); calcium-dependent exocytosis; Computational and Systems Biology; Exocytosis; mathematical modeling; Membrane vesicles; Neuroscience; Neurotransmission; Neurotransmitter release; Phospholipids; Plasma; synaptic transmission; Synaptic vesicles; Synaptotagmin; Vesicle fusion
• ISSN: 2050-084X; 2050-084X
• DOI: 10.7554/eLife.74810

Synaptic communication relies on the fusion of synaptic vesicles with the plasma membrane, which leads to neurotransmitter release. This exocytosis is triggered by brief and local elevations of intracellular Ca 2+ with remarkably high sensitivity. How this is molecularly achieved is unknown. While synaptotagmins confer the Ca 2+ sensitivity of neurotransmitter exocytosis, biochemical measurements reported Ca 2+ affinities too low to account for synaptic function. However, synaptotagmin’s Ca 2+ affinity increases upon binding the plasma membrane phospholipid PI(4,5)P 2 and, vice versa, Ca 2+ binding increases synaptotagmin’s PI(4,5)P 2 affinity, indicating a stabilization of the Ca 2+ /PI(4,5)P 2 dual-bound state. Here, we devise a molecular exocytosis model based on this positive allosteric stabilization and the assumptions that (1.) synaptotagmin Ca 2+ /PI(4,5)P 2 dual binding lowers the energy barrier for vesicle fusion and that (2.) the effect of multiple synaptotagmins on the energy barrier is additive. The model, which relies on biochemically measured Ca 2+ /PI(4,5)P 2 affinities and protein copy numbers, reproduced the steep Ca 2+ dependency of neurotransmitter release. Our results indicate that each synaptotagmin engaging in Ca 2+ /PI(4,5)P 2 dual-binding lowers the energy barrier for vesicle fusion by ~5 k B T and that allosteric stabilization of this state enables the synchronized engagement of several (typically three) synaptotagmins for fast exocytosis. Furthermore, we show that mutations altering synaptotagmin’s allosteric properties may show dominant-negative effects, even though synaptotagmins act independently on the energy barrier, and that dynamic changes of local PI(4,5)P 2 (e.g. upon vesicle movement) dramatically impact synaptic responses. We conclude that allosterically stabilized Ca 2+ /PI(4,5)P 2 dual binding enables synaptotagmins to exert their coordinated function in neurotransmission. For our brains and nervous systems to work properly, the nerve cells within them must be able to ‘talk’ to each other. They do this by releasing chemical signals called neurotransmitters which other cells can detect and respond to. Neurotransmitters are packaged in tiny membrane-bound spheres called vesicles. When a cell of the nervous system needs to send a signal to its neighbours, the vesicles fuse with the outer membrane of the cell, discharging their chemical contents for other cells to detect. The initial trigger for neurotransmitter release i