Identification of a Munc13-sensitive step in chromaffin cell large dense-core vesicle exocytosis

eLife

Volumn 4, Issue NOVEMBER2015, 2015, Pages

  Man, Kwun Nok M ^a   Imig, Cordelia ^a   Walter, Alexander M ^b   Pinheiro, Paulo S ^c   Stevens, David R ^d   Rettig, Jens ^d   Sørensen, Jakob B ^c   Cooper, Benjamin H ^a   Brose, Nils ^a   Wojcik, Sonja M ^a  

^a MAX PLANCK INSTITUTE OF EXPERIMENTAL MEDICINE   (Germany)

^b LEIBNIZ INSTITUT FÜR MOLEKULARE PHARMAKOLOGIE   (Germany)

^c UNIVERSITY OF COPENHAGEN   (Denmark)

^d SAARLAND UNIVERSITY   (Germany)

Author keywords

[No Author keywords available]

Indexed keywords

BAIAP3; CALCIUM ION; CATECHOLAMINE; CHROMOGRANIN A; GREEN FLUORESCENT PROTEIN; MUNC13; PROTEIN; SNARE PROTEIN; UNCLASSIFIED DRUG; YELLOW FLUORESCENT PROTEIN; NERVE PROTEIN; SIGNAL PEPTIDE; UNC13B PROTEIN, MOUSE;

ADULT; AMPEROMETRY; ANIMAL CELL; ANIMAL EXPERIMENT; ANIMAL MODEL; ANIMAL TISSUE; ARTICLE; CALCIUM CELL LEVEL; CHROMAFFIN CELL; CONTROLLED STUDY; CYTOPLASM VESICLE; ELECTRON TOMOGRAPHY; EMBRYO; EXOCYTOSIS; GENE; LARGE DENSE CORE VESICLE; MOLECULAR DOCKING; MOUSE; NONHUMAN; PROTEIN EXPRESSION; SYNAPSE VESICLE; UNC13B GENE; WESTERN BLOTTING; WHOLE CELL PATCH CLAMP; ANIMAL; BIOLOGICAL MODEL; METABOLISM; SECRETORY VESICLE;

ANIMALS; CHROMAFFIN CELLS; EXOCYTOSIS; INTRACELLULAR SIGNALING PEPTIDES AND PROTEINS; MICE; MODELS, BIOLOGICAL; NERVE TISSUE PROTEINS; SECRETORY VESICLES;

EID: 84957928455     PISSN: None     EISSN: 2050084X     Source Type: Journal    
DOI: 10.7554/elife.10635     Document Type: Article

References (85)

1. Ashery, U., A. Betz, T. Xu, N. Brose, and J. Rettig. 1999. An efficient method for infection of adrenal chromaffin cells using the Semliki Forest virus gene expression system. Eur. J. Cell Biol. 78:525-532.
2. Ashery, U., F. Varoqueaux, T. Voets, A. Betz, P. Thakur, H. Koch, E. Neher, N. Brose, and J. Rettig. 2000. Munc13-1 acts as a priming factor for large dense947 core vesicles in bovine chromaffin cells. EMBO J. 19:3586-3596.
3. Augustin, I., S. Korte, M. Rickmann, H.A. Kretzschmar, T.C. Sudhof, J.W. Herms, and N. Brose. 2001. The cerebellum-specific Munc13 isoform Munc13-3 regulates cerebellar synaptic transmission and motor learning in mice. J. Neurosci. 21:10-17.
4. Augustin, I., C. Rosenmund, T.C. Sudhof, and N. Brose. 1999. Munc13-1 is essential for fusion competence of glutamatergic synaptic vesicles. Nature. 400:457-461.
5. Bauer, C.S., R.J. Woolley, A.G. Teschemacher, and E.P. Seward. 2007. Potentiation of exocytosis by phospholipase C-coupled G-protein-coupled receptors requires the priming protein Munc13-1. J. Neurosci. 27:212-219.
6. Borisovska, M., Y. Zhao, Y. Tsytsyura, N. Glyvuk, S. Takamori, U. Matti, J. Rettig, T. Sudhof, and D. Bruns. 2005. v-SNAREs control exocytosis of vesicles from priming to fusion. EMBO J. 24:2114-2126.
7. Boswell, K.L., D.J. James, J.M. Esquibel, S. Bruinsma, R. Shirakawa, H. Horiuchi, and T.F. Martin. 2012. Munc13-4 reconstitutes calcium-dependent SNARE963 mediated membrane fusion. J. Cell Biol. 197:301-312.
8. Bruns, D. 2004. Detection of transmitter release with carbon fiber electrodes. Methods. 33:312-321.
9. Cooper, B., M. Hemmerlein, J. Ammermuller, C. Imig, K. Reim, N. Lipstein, S. Kalla, H. Kawabe, N. Brose, J.H. Brandstatter, and F. Varoqueaux. 2012. Munc13-independent vesicle priming at mouse photoreceptor ribbon synapses. J. Neurosci. 32:8040-8052.
10. Crozat, K., K. Hoebe, S. Ugolini, N.A. Hong, E. Janssen, S. Rutschmann, S. Mudd, S. Sovath, E. Vivier, and B. Beutler. 2007. Jinx, an MCMV susceptibility phenotype caused by disruption of Unc13d: a mouse model of type 3 familial hemophagocytic lymphohistiocytosis. J. Exp. Med. 204:853-863.
11. Daily, N.J., K.L. Boswell, D.J. James, and T.F. Martin. 2010. Novel interactions of CAPS (Ca2+-dependent activator protein for secretion) with the three neuronal SNARE proteins required for vesicle fusion. J. Biol. Chem. 977 285:35320-35329.
12. de Wit, H. 2010. Molecular mechanism of secretory vesicle docking. Biochem. Soc. 979 Trans. 38:192-198.
13. de Wit, H., L.N. Cornelisse, R.F. Toonen, and M. Verhage. 2006. Docking of secretory vesicles is syntaxin dependent. PLoS One. 1:e126.
14. de Wit, H., A.M. Walter, I. Milosevic, A. Gulyas-Kovacs, D. Riedel, J.B. Sorensen, and M. Verhage. 2009. Synaptotagmin-1 docks secretory vesicles to syntaxin-1/SNAP-25 acceptor complexes. Cell. 138:935-946.
15. Elhamdani, A., T.F. Martin, J.A. Kowalchyk, and C.R. Artalejo. 1999. Ca(2+)-dependent activator protein for secretion is critical for the fusion of dense-core vesicles with the membrane in calf adrenal chromaffin cells. J. Neurosci. 19:7375-7383.
16. Fasshauer, D., and M. Margittai. 2004. A transient N-terminal interaction of SNAP-25 and syntaxin nucleates SNARE assembly. J. Biol. Chem. 279:7613-7621.
17. Feldmann, J., I. Callebaut, G. Raposo, S. Certain, D. Bacq, C. Dumont, N. Lambert, M. Ouachee-Chardin, G. Chedeville, H. Tamary, V. Minard-Colin, E. Vilmer, S. Blanche, F. Le Deist, A. Fischer, and G. de Saint Basile. 2003. Munc13-4 is essential for cytolytic granules fusion and is mutated in a form of familial hemophagocytic lymphohistiocytosis (FHL3). Cell. 115:461-473.
18. Gandasi, N.R., and S. Barg. 2014. Contact-induced clustering of syntaxin and munc18 docks secretory granules at the exocytosis site. Nat. Commun. 5:3914.
19. Gao, Y., S. Zorman, G. Gundersen, Z. Xi, L. Ma, G. Sirinakis, J.E. Rothman, and Y. Zhang. 2012. Single reconstituted neuronal SNARE complexes zipper in three distinct stages. Science. 337:1340-1343.
20. Gerber, S.H., J.C. Rah, S.W. Min, X. Liu, H. de Wit, I. Dulubova, A.C. Meyer, J. Rizo, M. Arancillo, R.E. Hammer, M. Verhage, C. Rosenmund, and T.C. Sudhof. 2008. Conformational switch of syntaxin-1 controls synaptic vesicle fusion. Science. 321:1507-1510.
21. Graham, M.E., M.T. Handley, J.W. Barclay, L.F. Ciufo, S.L. Barrow, A. Morgan, and R.D. Burgoyne. 2008. A gain-of-function mutant of Munc18-1 stimulates secretory granule recruitment and exocytosis and reveals a direct interaction of Munc18-1 with Rab3. Biochem. J. 409:407-416.
22. Gulyas-Kovacs, A., H. de Wit, I. Milosevic, O. Kochubey, R. Toonen, J. Klingauf, M. Verhage, and J.B. Sorensen. 2007. Munc18-1: sequential interactions with the fusion machinery stimulate vesicle docking and priming. J. Neurosci. 27:8676-8686.
23. Hammarlund, M., M.T. Palfreyman, S. Watanabe, S. Olsen, and E.M. Jorgensen. 2007. Open syntaxin docks synaptic vesicles. PLoS Biol. 5:e198.
24. Hammarlund, M., S. Watanabe, K. Schuske, and E.M. Jorgensen. 2008. CAPS and syntaxin dock dense core vesicles to the plasma membrane in neurons. J. Cell Biol. 180:483-491.
25. Han, G.A., N.T. Malintan, N.M. Saw, L. Li, L. Han, F.A. Meunier, B.M. Collins, and S. Sugita. 2011. Munc18-1 domain-1 controls vesicle docking and secretion by interacting with syntaxin-1 and chaperoning it to the plasma membrane. Mol. Biol. Cell. 22:4134-4149.
26. Honigmann, A., G. van den Bogaart, E. Iraheta, H.J. Risselada, D. Milovanovic, V. Mueller, S. Mullar, U. Diederichsen, D. Fasshauer, H. Grubmuller, S.W. Hell, C. Eggeling, K. Kuhnel, and R. Jahn. 2013. Phosphatidylinositol 4,5-bisphosphate clusters act as molecular beacons for vesicle recruitment. Nat. Struct. Mol. Biol. 20:679-686.
27. Imig, C., S.W. Min, S. Krinner, M. Arancillo, C. Rosenmund, T.C. Sudhof, J.S. Rhee, N. Brose, and B.H. Cooper. 2014. The morphological and molecular nature of synpatic vesicle priming at presynaptic active zones. Neuron. 82:416-431.
28. James, D.J., and T.F. Martin. 2013. CAPS and Munc13: CATCHRs that SNARE Vesicles. Front. Endocrinol. (Lausanne). 4:187.
29. Jockusch, W.J., D. Speidel, A. Sigler, J.B. Sorensen, F. Varoqueaux, J.S. Rhee, and N. Brose. 2007. CAPS-1 and CAPS-2 are essential synaptic vesicle priming proteins. Cell. 131:796-808.
30. Kabachinski, G., M. Yamaga, D.M. Kielar-Grevstad, S. Bruinsma, and T.F. Martin. 2014. CAPS and Munc13 utilize distinct PIP2-linked mechanisms to promote vesicle exocytosis. Mol. Biol. Cell. 25:508-521.
31. Kalla, S., M. Stern, J. Basu, F. Varoqueaux, K. Reim, C. Rosenmund, N.E. Ziv, and N. Brose. 2006. Molecular dynamics of a presynaptic active zone protein studied in Munc13-1-enhanced yellow fluorescent protein knock-in mutant mice. J. Neurosci. 26:13054-13066.
32. Kang, L., Z. He, P. Xu, J. Fan, A. Betz, N. Brose, and T. Xu. 2006. Munc13-1 is required for the sustained release of insulin from pancreatic beta cells. Cell. Metab. 3:463-468.
33. Koch, H., K. Hofmann, and N. Brose. 2000. Definition of Munc13-homology-domains and characterization of a novel ubiquitously expressed Munc13 isoform. Biochem. J. 349:247-253.
34. Kremer, J.R., D.N. Mastronarde, and J.R. Mcintosh. 1996. Computer visualization of three-dimensional image data using IMOD. J. Struct. Biol. 116:71-76.
35. Kwan, E.P., L. Xie, L. Sheu, C.J. Nolan, M. Prentki, A. Betz, N. Brose, and H.Y. Gaisano. 2006. Munc13-1 deficiency reduces insulin secretion and causes abnormal glucose tolerance. Diabetes. 55:1421-1429.
36. Lecat, S., H.W. Matthes, R. Pepperkok, J.C. Simpson, and J.L. Galzi. 2015. A Fluorescent Live Imaging Screening Assay Based on Translocation Criteria Identifies Novel Cytoplasmic Proteins Implicated in G Protein-coupled Receptor Signaling Pathways. Mol. Cell. Proteomics. 14:1385-1399.
37. Liu, Y., C. Schirra, L. Edelmann, U. Matti, J. Rhee, D. Hof, D. Bruns, N. Brose, H. Rieger, D.R. Stevens, and J. Rettig. 2010. Two distinct secretory vesicle-priming steps in adrenal chromaffin cells. J. Cell Biol. 190:1067-1077.
38. Liu, Y., C. Schirra, D.R. Stevens, U. Matti, D. Speidel, D. Hof, D. Bruns, N. Brose, and J. Rettig. 2008. CAPS facilitates filling of the rapidly releasable pool of large dense-core vesicles. J. Neurosci. 28:5594-5601.
39. Ma, C., W. Li, Y. Xu, and J. Rizo. 2011. Munc13 mediates the transition from the closed syntaxin-Munc18 complex to the SNARE complex. Nat. Struct. Mol. Biol. 18:542-549.
40. Ma, C., L. Su, A.B. Seven, Y. Xu, and J. Rizo. 2013. Reconstitution of the vital functions of Munc18 and Munc13 in neurotransmitter release. Science. 339:421-425.
41. Mastronarde, D.N. 2005. Automated electron microscope tomography using robust prediction of specimen movements. J. Struct. Biol. 152:36-51.
42. Mobius, W., B. Cooper, W.A. Kaufmann, C. Imig, T. Ruhwedel, N. Snaidero, A.S. Saab, and F. Varoqueaux. 2010. Electron microscopy of the mouse central nervous system. Methods Cell Biol. 96:475-512.
43. Moser, T., and E. Neher. 1997. Rapid exocytosis in single chromaffin cells recorded from mouse adrenal slices. J. Neurosci. 17:2314-2323.
44. Mosharov, E.V., and D. Sulzer. 2005. Analysis of exocytotic events recorded by amperometry. Nat. Methods. 2:651-658.
45. Neher, E. 2006. A comparison between exocytic control mechanisms in adrenal chromaffin cells and a glutamatergic synapse. Pflugers Arch. 453:261-268.
46. Ovsepian, S.V., and J.O. Dolly. 2011. Dendritic SNAREs add a new twist to the old neuron theory. Proc. Natl. Acad. Sci. U. S. A. 108:19113-19120.
47. Pang, Z.P., and T.C. Sudhof. 2010. Cell biology of Ca2+-triggered exocytosis. Curr. Opin. Cell Biol. 22:496-505.
48. Parisotto, D., J. Malsam, A. Scheutzow, J.M. Krause, and T.H. Sollner. 2012. SNAREpin assembly by Munc18-1 requires previous vesicle docking by synaptotagmin 1. J. Biol. Chem. 287:31041-31049.
49. Park, Y., J.B. Seo, A. Fraind, A. Perez-Lara, H. Yavuz, K. Han, S.R. Jung, I. Kattan, P.J. Walla, M. Choi, D.S. Cafiso, D.S. Koh, and R. Jahn. 2015. Synaptotagmin-1 binds to PIP2-containing membrane but not to SNAREs at physiological ionic strength. Nat. Struct. Mol. Biol. 22:815-823.
50. Parsons, T.D., J.R. Coorssen, H. Horstmann, and W. Almers. 1995. Docked granules, the exocytic burst, and the need for ATP hydrolysis in endocrine cells. Neuron. 15:1085-1096.
51. Pinheiro, P.S., A.M. Jansen, H. de Wit, B. Tawfik, K.L. Madsen, M. Verhage, U. Gether, and J.B. Sorensen. 2014. The BAR domain protein PICK1 controls vesicle number and size in adrenal chromaffin cells. J. Neurosci. 34:1068810700.
52. Plattner, H., A.R. Artalejo, and E. Neher. 1997. Ultrastructural organization of bovine chromaffin cell cortex-analysis by cryofixation and morphometry of aspects pertinent to exocytosis. J. Cell Biol. 139:1709-1717.
53. Pobbati, A.V., A. Stein, and D. Fasshauer. 2006. N- to C-terminal SNARE complex assembly promotes rapid membrane fusion. Science. 313:673-676.
54. Renden, R., B. Berwin, W. Davis, K. Ann, C.T. Chin, R. Kreber, B. Ganetzky, T.F. Martin, and K. Broadie. 2001. Drosophila CAPS is an essential gene that regulates dense-core vesicle release and synaptic vesicle fusion. Neuron. 31:421-437.
55. Richmond, J.E., W.S. Davis, and E.M. Jorgensen. 1999. UNC-13 is required for synaptic vesicle fusion in C. elegans. Nat. Neurosci. 2:959-964.
56. Rosenmund, C., A. Sigler, I. Augustin, K. Reim, N. Brose, and J.S. Rhee. 2002. Differential control of vesicle priming and short-term plasticity by Munc13 isoforms. Neuron. 33:411-424.
57. Rostaing, P., E. Real, L. Siksou, J.P. Lechaire, T. Boudier, T.M. Boeckers, F. Gertler, E.D. Gundelfinger, A. Triller, and S. Marty. 2006. Analysis of synaptic ultrastructure without fixative using high-pressure freezing and tomography. Eur. J. Neurosci. 24:3463-3474.
58. Sassa, T., S. Harada, H. Ogawa, J.B. Rand, I.N. Maruyama, and R. Hosono. 1999. Regulation of the UNC-18-Caenorhabditis elegans syntaxin complex by UNC-13. J. Neurosci. 19:4772-4777.
59. Schonn, J.S., J.R. van Weering, R. Mohrmann, O.M. Schluter, T.C. Sudhof, H. de Wit, M. Verhage, and J.B. Sorensen. 2010. Rab3 proteins involved in vesicle biogenesis and priming in embryonic mouse chromaffin cells. Traffic. 11:1415-1428.
60. Shirakawa, R., T. Higashi, A. Tabuchi, A. Yoshioka, H. Nishioka, M. Fukuda, T. Kita, and H. Horiuchi. 2004. Munc13-4 is a GTP-Rab27-binding protein regulating dense core granule secretion in platelets. J. Biol. Chem. 279:1073010737.
61. Siksou, L., F. Varoqueaux, O. Pascual, A. Triller, N. Brose, and S. Marty. 2009. A common molecular basis for membrane docking and functional priming of synaptic vesicles. Eur. J. Neurosci. 30:49-56.
62. Sorensen, J.B., R. Fernandez-Chacon, T.C. Sudhof, and E. Neher. 2003a. Examining synaptotagmin 1 function in dense core vesicle exocytosis under direct control of Ca2+. J. Gen. Physiol. 122:265-276.
63. Sorensen, J.B., G. Nagy, F. Varoqueaux, R.B. Nehring, N. Brose, M.C. Wilson, and E. Neher. 2003b. Differential control of the releasable vesicle pools by SNAP-25 splice variants and SNAP-23. Cell. 114:75-86.
64. Sorensen, J.B., K. Wiederhold, E.M. Muller, I. Milosevic, G. Nagy, B.L. de Groot, H. Grubmuller, and D. Fasshauer. 2006. Sequential N- to C-terminal SNARE complex assembly drives priming and fusion of secretory vesicles. EMBO J. 25:955-966.
65. Speese, S., M. Petrie, K. Schuske, M. Ailion, K. Ann, K. Iwasaki, E.M. Jorgensen, and T.F. Martin. 2007. UNC-31 (CAPS) is required for dense-core vesicle but not synaptic vesicle exocytosis in Caenorhabditis elegans. J. Neurosci. 27:6150-6162.
66. Speidel, D., A. Salehi, S. Obermueller, I. Lundquist, N. Brose, E. Renstrom, and P. Rorsman. 2008. CAPS1 and CAPS2 regulate stability and recruitment of insulin granules in mouse pancreatic beta cells. Cell. Metab. 7:57-67.
67. Stevens, D.R., Z.X. Wu, U. Matti, H.J. Junge, C. Schirra, U. Becherer, S.M. Wojcik, N. Brose, and J. Rettig. 2005. Identification of the minimal protein domain required for priming activity of Munc13-1. Curr. Biol. 15:2243-2248.
68. Toonen, R.F., O. Kochubey, H. de Wit, A. Gulyas-Kovacs, B. Konijnenburg, J.B. Sorensen, J. Klingauf, and M. Verhage. 2006. Dissecting docking and tethering of secretory vesicles at the target membrane. EMBO J. 25:37253737.
69. Tsuboi, T., and M. Fukuda. 2006. Rab3A and Rab27A cooperatively regulate the docking step of dense-core vesicle exocytosis in PC12 cells. J. Cell Sci. 119:2196-2203.
70. van de Bospoort, R., M. Farina, S.K. Schmitz, A. de Jong, H. de Wit, M. Verhage, and R.F. Toonen. 2012. Munc13 controls the location and efficiency of dense-core vesicle release in neurons. J. Cell Biol. 199:883-891.
71. van Weering, J.R., R.F. Toonen, and M. Verhage. 2007. The role of Rab3a in secretory vesicle docking requires association/dissociation of guanidine phosphates and Munc18-1. PLoS One. 2:e616.
72. Varoqueaux, F., A. Sigler, J.S. Rhee, N. Brose, C. Enk, K. Reim, and C. Rosenmund. 2002. Total arrest of spontaneous and evoked synaptic transmission but normal synaptogenesis in the absence of Munc13-mediated vesicle priming. Proc. Natl. Acad. Sci. U. S. A. 99:9037-9042.
73. Varoqueaux, F., M.S. Sons, J.J. Plomp, and N. Brose. 2005. Aberrant morphology and residual transmitter release at the Munc13-deficient mouse neuromuscular synapse. Mol. Cell. Biol. 25:5973-5984.
74. Voets, T. 2000. Dissection of three Ca2+-dependent steps leading to secretion in chromaffin cells from mouse adrenal slices. Neuron. 28:537-545.
75. Voets, T., E. Neher, and T. Moser. 1999. Mechanisms underlying phasic and sustained secretion in chromaffin cells from mouse adrenal slices. Neuron. 23:607-615.
76. Voets, T., R.F. Toonen, E.C. Brian, H. de Wit, T. Moser, J. Rettig, T.C. Sudhof, E. Neher, and M. Verhage. 2001. Munc18-1 promotes large dense-core vesicle docking. Neuron. 31:581-591.
77. Walter, A.M., P.S. Pinheiro, M. Verhage, and J.B. Sorensen. 2013. A sequential vesicle pool model with a single release sensor and a Ca(2+)-dependent priming catalyst effectively explains Ca(2+)-dependent properties of neurosecretion. PLoS Comput. Biol. 9:e1003362.
78. Walter, A.M., K. Wiederhold, D. Bruns, D. Fasshauer, and J.B. Sorensen. 2010. Synaptobrevin N-terminally bound to syntaxin-SNAP-25 defines the primed vesicle state in regulated exocytosis. J. Cell Biol. 188:401-413.
79. Weimer, R.M., E.O. Gracheva, O. Meyrignac, K.G. Miller, J.E. Richmond, and J.L. Bessereau. 2006. UNC-13 and UNC-10/rim localize synaptic vesicles to specific membrane domains. J. Neurosci. 26:8040-8047.
80. Wojcik, S.M., M. Tantra, B. Stepniak, K.N. Man, K. Muller-Ribbe, M. Begemann, A. Ju, S. Papiol, A. Ronnenberg, A. Gurvich, Y. Shin, I. Augustin, N. Brose, and H. Ehrenreich. 2013. Genetic markers of a Munc13 protein family member, BAIAP3, are gender specifically associated with anxiety and benzodiazepine abuse in mice and humans. Mol. Med. 19:135-148.
81. Wu, Y., Y. Gu, M.K. Morphew, J. Yao, F.L. Yeh, M. Dong, and E.R. Chapman. 2012. All three components of the neuronal SNARE complex contribute to secretory vesicle docking. J. Cell Biol. 198:323-330.
82. Yang, X., S. Wang, Y. Sheng, M. Zhang, W. Zou, L. Wu, L. Kang, J. Rizo, R. Zhang, T. Xu, and C. Ma. 2015. Syntaxin opening by the MUN domain underlies the function of Munc13 in synaptic-vesicle priming. Nat. Struct. Mol. Biol. 22:547-554.
83. Zhou, K.M., Y.M. Dong, Q. Ge, D. Zhu, W. Zhou, X.G. Lin, T. Liang, Z.X. Wu, and T. Xu. 2007. PKA activation bypasses the requirement for UNC-31 in the docking of dense core vesicles from C. elegans neurons. Neuron. 56:657-669.
84. Zhou, Z., and S. Misler. 1995. Action potential-induced quantal secretion of catecholamines from rat adrenal chromaffin cells. J. Biol. Chem. 270:34983505.
85. Zikich, D., A. Mezer, F. Varoqueaux, A. Sheinin, H.J. Junge, E. Nachliel, R. Melamed, N. Brose, M. Gutman, and U. Ashery. 2008. Vesicle priming and recruitment by ubMunc13-2 are differentially regulated by calcium and calmodulin. J. Neurosci. 28:1949-1960.