Application Notes

Interactions of liposome embedded SNARE proteins

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Protein-Vesicle Interaction Analysis Application Note NT-MO-002 Interactions of Liposome embedded SNARE Proteins Karsten Meyenberg 1 and Geert van den Bogaart 2 1 Institut für Organische und Biomolekulare Chemie, Georg-August-Universität Göttingen, Germany 2 Max-Planck-Institut für biophysikalische Chemie, Göttingen, Germany Abstract SNARE protein complexes play a key role in membrane fusion processes, e.g. the fusion of synaptic vesicles with the pre-synaptic plasma membrane in response to elevated Ca 2+ levels. This study aimed to demonstrate the use of MicroScale Thermophoresis for the detection of protein-protein interaction in more complex assay systems like in this case the interaction of membrane proteins embedded into liposomes. Introduction Synaptical signal transduction to neighboring neuronal cells occurs mainly by chemical signal transduction. Fig. 1 Molecular modell of a synaptic vesicle (Reinhard Jahn, Max-Planck-Institut für biophysikalische Chemie, Göttingen, Germany) Neurotransmitters are released from the pre- synaptic cell inducing activation of ion-channels which in turn induces an electrical signal in the post-synaptical cell. The neurotransmitters are stored in multiple pre-synaptic vesicles closely attached to the plasma membrane to ensure a fast release upon incoming signals. Voltage-gated Ion channels lead to elevated Ca 2+ levels in the pre- synaptic cell and induce fusion of the synaptic vesicles with the plasma membrane with the subsequent release of the neurotransmitters into the synaptic cleft. Fig. 2 The SNARE-complex attaches vesicles to the cell membrane. Three proteins – syntaxin (red), synaptobrevin-2 (syp-2) (blue) and SNAP25 (green) – are forming a helical complex. Syntaxin and synaptobrevin-2 are integral membrane proteins bringing vesicle and cell membrane in close contact. (Reinhard Jahn, Max-Planck-Institut für biophysikalische Chemie, Göttingen, Germany) In eukaryotes, most intracellular membrane fusion reactions are mediated by the interaction of complementary SNARE proteins that are present in both fusing membranes. SNARE complexes consist of SNAP25 and the membrane protein syntaxin which is embedded in the plasma

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