Calcium Promotes the Formation of Syntaxin 1 Mesoscale Domains through Phosphatidylinositol 4,5-Bisphosphate

Phosphatidylinositol 4,5-bisphosphate (PI(4,5)P2) is a minor component of total plasma membrane lipids, but it has a substantial role in the regulation of many cellular functions, including exo- and endocytosis. Recently, it was shown that PI(4,5)P2 and syntaxin 1, a SNARE protein that catalyzes regulated exocytosis, form domains in the plasma membrane that constitute recognition sites for vesicle docking. Also, calcium was shown to promote syntaxin 1 clustering in the plasma membrane, but the molecular mechanism was unknown. Here, using a combination of superresolution stimulated emission depletion microscopy, FRET, and atomic force microscopy, we show that Ca2+ acts as a charge bridge that specifically and reversibly connects multiple syntaxin 1/PI(4,5)P2 complexes into larger mesoscale domains. This transient reorganization of the plasma membrane by physiological Ca2+ concentrations is likely to be important for Ca2+-regulated secretion.

[1]  R. Jahn,et al.  Organization and dynamics of SNARE proteins in the presynaptic membrane , 2015, Front. Physiol..

[2]  S. Hell,et al.  Hydrophobic mismatch sorts SNARE proteins into distinct membrane domains , 2015, Nature Communications.

[3]  Nikhil R. Gandasi,et al.  Contact-induced clustering of syntaxin and munc18 docks secretory granules at the exocytosis site , 2014, Nature Communications.

[4]  S. Rizzoli.  Synaptic vesicle recycling: steps and principles , 2014, The EMBO journal.

[5]  M. Prieto,et al.  Ca(2+) induces PI(4,5)P2 clusters on lipid bilayers at physiological PI(4,5)P2 and Ca(2+) concentrations. , 2014, Biochimica et biophysica acta.

[6]  T. Südhof,et al.  Neurotransmitter Release: The Last Millisecond in the Life of a Synaptic Vesicle , 2013, Neuron.

[7]  Alf Honigmann,et al.  Coaligned dual-channel STED nanoscopy and molecular diffusion analysis at 20 nm resolution. , 2013, Biophysical journal.

[8]  S. Hell,et al.  Phosphatidylinositol 4,5-bisphosphate clusters act as molecular beacons for vesicle recruitment , 2013, Nature Structural &Molecular Biology.

[9]  R. Habets,et al.  Synaptic PI(3,4,5)P3 Is Required for Syntaxin1A Clustering and Neurotransmitter Release , 2013, Neuron.

[10]  G. van den Bogaart,et al.  Microdomains of SNARE proteins in the plasma membrane. , 2013, Current topics in membranes.

[11]  R. Jahn,et al.  Molecular machines governing exocytosis of synaptic vesicles , 2012, Nature.

[12]  G. van den Bogaart,et al.  Controlling synaptotagmin activity by electrostatic screening , 2012, Nature Structural &Molecular Biology.

[13]  Johannes E. Schindelin,et al.  Fiji: an open-source platform for biological-image analysis , 2012, Nature Methods.

[14]  Mike Heilemann,et al.  Super-resolution Imaging Reveals the Internal Architecture of Nano-sized Syntaxin Clusters* , 2012, The Journal of Biological Chemistry.

[15]  J. Carrascosa,et al.  Minimizing tip-sample forces in jumping mode atomic force microscopy in liquid. , 2012, Ultramicroscopy.

[16]  P. Janmey,et al.  Divalent cation-induced cluster formation by polyphosphoinositides in model membranes. , 2012, Journal of the American Chemical Society.

[17]  P. Janmey,et al.  Phosphatidylinositol 4, 5 bisphosphate and the actin cytoskeleton. , 2012, Sub-cellular biochemistry.

[18]  P. Lory,et al.  A Cav 3 . 2 / Syntaxin-1 A Signaling Complex Controls T-type Channel Activity and Low-threshold Exocytosis * , 2012 .

[19]  Andrea J Liu,et al.  Divalent cation-dependent formation of electrostatic PIP2 clusters in lipid monolayers. , 2011, Biophysical journal.

[20]  L. Tamm,et al.  Molecular mechanism of cholesterol- and polyphosphoinositide-mediated syntaxin clustering. , 2011, Biochemistry.

[21]  Hayder Amin,et al.  Membrane protein sequestering by ionic protein-lipid interactions , 2011, Nature.

[22]  J. N. Davies,et al.  Bipartite syntaxin 1A interactions mediate CaV2.2 calcium channel regulation. , 2011, Biochemical and biophysical research communications.

[23]  R. Jahn,et al.  Ca2+ induces clustering of membrane proteins in the plasma membrane via electrostatic interactions , 2011, The EMBO journal.

[24]  Amparo Gil,et al.  The F-actin cortical network is a major factor influencing the organization of the secretory machinery in chromaffin cells , 2011, Journal of Cell Science.

[25]  Michelle K. Knowles,et al.  Syntaxin clusters assemble reversibly at sites of secretory granules in live cells , 2010, Proceedings of the National Academy of Sciences.

[26]  O. Mandula,et al.  t-SNARE Protein Conformations Patterned by the Lipid Microenvironment* , 2010, The Journal of Biological Chemistry.

[27]  P. Lappalainen,et al.  Regulation of the actin cytoskeleton-plasma membrane interplay by phosphoinositides. , 2010, Physiological reviews.

[28]  J. Pessin,et al.  Evidence that electrostatic interactions between vesicle-associated membrane protein 2 and acidic phospholipids may modulate the fusion of transport vesicles with the plasma membrane. , 2009, Molecular biology of the cell.

[29]  P. Janmey,et al.  Calcium-dependent lateral organization in phosphatidylinositol 4,5-bisphosphate (PIP2)- and cholesterol-containing monolayers. , 2009, Biochemistry.

[30]  Catherine Picart,et al.  Giant unilamellar vesicles containing phosphatidylinositol(4,5)bisphosphate: characterization and functionality. , 2008, Biophysical journal.

[31]  Edwin R Chapman,et al.  How does synaptotagmin trigger neurotransmitter release? , 2008, Annual review of biochemistry.

[32]  Thorsten Lang,et al.  Anatomy and Dynamics of a Supramolecular Membrane Protein Cluster , 2007, Science.

[33]  B. Davletov,et al.  Tight coupling of the t-SNARE and calcium channel microdomains in adrenomedullary slices and not in cultured chromaffin cells. , 2007, Cell calcium.

[34]  Thorsten Lang,et al.  The SNARE motif is essential for the formation of syntaxin clusters in the plasma membrane. , 2006, Biophysical journal.

[35]  E. Neher,et al.  Presynaptic calcium and control of vesicle fusion , 2005, Current Opinion in Neurobiology.

[36]  S. Terakawa,et al.  The Activation of Exocytotic Sites by the Formation of Phosphatidylinositol 4,5-Bisphosphate Microdomains at Syntaxin Clusters* , 2005, Journal of Biological Chemistry.

[37]  C. Schmidt,et al.  Resolving the molecular structure of microtubules under physiological conditions with scanning force microscopy , 2004, European Biophysics Journal.

[38]  D. Bruns,et al.  SNAREs are concentrated in cholesterol‐dependent clusters that define docking and fusion sites for exocytosis , 2001, The EMBO journal.

[39]  H. Thoenen,et al.  Relationship between NGF-mediated volume increase and "priming effect" in fast and slow reacting clones of PC12 pheochromocytoma cells. Role of cAMP. , 1983, Experimental cell research.

[40]  R. Weingart,et al.  Free magnesium in sheep, ferret and frog striated muscle at rest measured with ion‐selective micro‐electrodes , 1982, The Journal of physiology.

[41]  T. Reese.  Synaptic vesicle exocytosis. , 1981, JAMA.

[42]  E. Newsholme,et al.  The contents of adenine nucleotides, phosphagens and some glycolytic intermediates in resting muscles from vertebrates and invertebrates. , 1975, The Biochemical journal.