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Cited by 70Cellular transport and secretionPancreatic function and diabetesExtracellular vesicles in disease

SNARE Conformational Changes that Prepare Vesicles for Exocytosis

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Takahashi, Noriko, Hatakeyama, Hiroyasu, Okado, Haruo, Noguchi, Jun, Ohno, Mitsuyo, Kasai, Haruo·2010·Cell Metabolism
DOI 10.1016/j.cmet.2010.05.013
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Abstract

When cells release hormones and neurotransmitters through exocytosis, cytosolic Ca(2+) triggers the fusion of secretory vesicles with the plasma membrane. It is well known that this fusion requires assembly of a SNARE protein complex. However, the timing of SNARE assembly relative to vesicle fusion--essential for understanding exocytosis--has not been demonstrated. To investigate this timing, we constructed a probe that detects the assembly of two plasma membrane SNAREs, SNAP25 and syntaxin-1A, through fluorescence resonance energy transfer (FRET). With two-photon imaging, we simultaneously measured FRET signals and insulin exocytosis in beta cells from the pancreatic islet of Langerhans. In some regions of the cell, we found that the SNARE complex was preassembled, which enabled rapid exocytosis. In other regions, SNARE assembly followed Ca(2+) influx, and exocytosis was slower. Thus, SNARE proteins exist in multiple stable preparatory configurations, from which Ca(2+) may trigger exocytosis through distinct mechanisms and with distinct kinetics.

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  • Using a FRET probe that reports the conformational state of the t-SNARE SNAP25, the SNARE complex exists in two different conformations: a tight (high-FRET) and a loose (low-FRET) conformation.

    “Using a fluorescence resonance energy transfer (FRET) probe that can report the conformational state of the t-SNARE SNAP25, it was shown that the SNARE complex exists in two different conformations: a tight (high-FRET) and a loose (low-FRET) conformation (661).”

    From Pancreatic β-Cell Electrical Activity and Insulin Secretion: Of Mice and Men (2018) · cites this paper as [661]

    The Results (page unavailable) describe a SNAP25 FRET probe and distinct high- and low-FRET populations. But the authors interpret these as SNAP25 assembly with syntaxin being complete or absent, not as tight and loose conformations of an existing SNARE complex. (LLM, conf=0.87)

    Experimental — accuracy not yet measured

  • In the low-FRET state, there was a significant delay (of up to 10 s) between elevation of [Ca2+]i and exocytosis, and an increase in FRET preceded fusion indicating the complex underwent a conformational change.

    “Conversely, in the low-FRET state, there was a significant delay (of up to 10 s) between elevation of [Ca2+]i and exocytosis, and an increase in FRET preceded fusion indicating the complex underwent a conformational change.”

    From Pancreatic β-Cell Electrical Activity and Insulin Secretion: Of Mice and Men (2018) · cites this paper as [661]

  • The high-FRET state was associated with rapid exocytosis.

    “The high-FRET state was associated with rapid exocytosis (visualised by Alexa 594 uptake into the granule lumen; see Figure 18D-E).”

    From Pancreatic β-Cell Electrical Activity and Insulin Secretion: Of Mice and Men (2018) · cites this paper as [661]

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Research works printed but not linked: 33

  1. [1]1. Allersma, M.W. ∙ Wang, L. ∙ Axelrod, D. ... Visualization of regulated exocytosis with a granule-membrane probe using total internal reflection microscopy Mol. Biol. Cell. 2004; 15:4658-4668 Crossref Scopus (111) PubMed Google Scholar
  2. [2]2. Alvarez de Toledo, G. ∙ Fernandez, J.M. Compound versus multigranular exocytosis in peritoneal mast cells J. Gen. Physiol. 1990; 95:397-409 Crossref Scopus (121) PubMed Google Scholar
  3. [3]3. An, S.J. ∙ Almers, W. Tracking SNARE complex formation in live endocrine cells Science. 2004; 306:1042-1046 Crossref Scopus (117) PubMed Google Scholar
  4. [4]4. Augustine, G.J. How does calcium trigger neurotransmitter release? Curr. Opin. Neurobiol. 2001; 11:320-326 Crossref Scopus (319) PubMed Google Scholar
  5. [5]5. Bhalla, A. ∙ Chicka, M.C. ∙ Tucker, W.C. ... Ca(2+)-synaptotagmin directly regulates t-SNARE function during reconstituted membrane fusion Nat. Struct. Mol. Biol. 2006; 13:323-330 Crossref Scopus (148) PubMed Google Scholar
  6. [6]6. Brunger, A.T. Structure and function of SNARE and SNARE-interacting proteins Q. Rev. Biophys. 2005; 38:1-47 Crossref Scopus (223) PubMed Google Scholar
  7. [7]7. Brunger, A.T. ∙ Weninger, K. ∙ Bowen, M. ... Single-molecule studies of the neuronal SNARE fusion machinery Annu. Rev. Biochem. 2009; 78:903-928 Crossref Scopus (89) PubMed Google Scholar
  8. [8]8. Bruns, D. ∙ Jahn, R. Real-time measurement of transmitter release from single synaptic vesicles Nature. 1995; 377:62-65 Crossref Scopus (374) PubMed Google Scholar
  9. [9]9. Chapman, E.R. How does synaptotagmin trigger neurotransmitter release? Annu. Rev. Biochem. 2008; 77:615-641 Crossref Scopus (437) PubMed Google Scholar
  10. [10]10. Chapman, E.R. ∙ Hanson, P.I. ∙ An, S. ... Ca2+ regulates the interaction between synaptotagmin and syntaxin 1 J. Biol. Chem. 1995; 270:23667-23671 Crossref Scopus (323) PubMed Google Scholar
  11. [11]11. Chicka, M.C. ∙ Hui, E. ∙ Liu, H. ... Synaptotagmin arrests the SNARE complex before triggering fast, efficient membrane fusion in response to Ca2+ Nat. Struct. Mol. Biol. 2008; 15:827-835 Crossref Scopus (161) PubMed Google Scholar
  12. [12]12. de Wit, H. ∙ Walter, A.M. ∙ Milosevic, I. ... Synaptotagmin-1 docks secretory vesicles to syntaxin-1/SNAP-25 acceptor complexes Cell. 2009; 138:935-946 Full Text Full Text (PDF) Scopus (223) PubMed Google Scholar
  13. [13]13. Fasshauer, D. ∙ Margittai, M. A transient N-terminal interaction of SNAP-25 and syntaxin nucleates SNARE assembly J. Biol. Chem. 2004; 279:7613-7621 Crossref Scopus (144) PubMed Google Scholar
  14. [14]14. Fasshauer, D. ∙ Antonin, W. ∙ Subramaniam, V. ... SNARE assembly and disassembly exhibit a pronounced hysteresis Nat. Struct. Biol. 2002; 9:144-151 Crossref Scopus (130) PubMed Google Scholar
  15. [15]15. Furuya, S. ∙ Edwards, C. ∙ Ornberg, R.L. Exocytosis of bovine chromaffin granules in Ficoll captured by rapid freezing J. Electron Microsc. (Tokyo). 1989; 38:143-147 PubMed Google Scholar
  16. [17]17. Hafez, I. ∙ Stolpe, A. ∙ Lindau, M. Compound exocytosis and cumulative fusion in eosinophils J. Biol. Chem. 2003; 278:44921-44928 Crossref Scopus (64) PubMed Google Scholar
  17. [18]18. Hatakeyama, H. ∙ Kishimoto, T. ∙ Nemoto, T. ... Rapid glucose sensing by protein kinase A for insulin exocytosis in mouse pancreatic islets J. Physiol. 2006; 570:271-282 Crossref Scopus (65) PubMed Google Scholar
  18. [19]19. Hatakeyama, H. ∙ Takahashi, N. ∙ Kishimoto, T. ... Two cAMP-dependent pathways differentially regulate exocytosis of large dense-core and small vesicles in mouse beta-cells J. Physiol. 2007; 582:1087-1098 Crossref Scopus (58) PubMed Google Scholar
  19. [20]20. Henquin, J.C. Regulation of insulin secretion: a matter of phase control and amplitude modulation Diabetologia. 2009; 52:739-751 Crossref Scopus (389) PubMed Google Scholar
  20. [21]21. Ichikawa, A. Fine structural changes in response to hormonal stimulation of the perfused canine pancreas J. Cell Biol. 1965; 24:369-385 Crossref Scopus (151) PubMed Google Scholar
  21. [22]22. Jahn, R. ∙ Südhof, T.C. Synaptic vesicles and exocytosis Annu. Rev. Neurosci. 1994; 17:219-246 Crossref Scopus (358) PubMed Google Scholar
  22. [23]23. Jahn, R. ∙ Lang, T. ∙ Südhof, T.C. Membrane fusion Cell. 2003; 112:519-533 Full Text Full Text (PDF) Scopus (1280) PubMed Google Scholar
  23. [24]24. Kahn, C.R. Joslin's Diabetes Melitus Lippincott Williams and Wilkins, Philadelphia, 2004 Google Scholar
  24. [25]25. Kasai, H. Comparative biology of Ca2+-dependent exocytosis: implications of kinetic diversity for secretory function Trends Neurosci. 1999; 22:88-93 Full Text Full Text (PDF) Scopus (167) PubMed Google Scholar
  25. [27]27. Kasai, H. ∙ Hatakeyama, H. ∙ Kishimoto, T. ... A new quantitative (two-photon extracellular polar-tracer imaging-based quantification (TEPIQ)) analysis for diameters of exocytic vesicles and its application to mouse pancreatic islets J. Physiol. 2005; 568:891-903 Crossref Scopus (27) PubMed Google Scholar
  26. [28]28. Kasai, H. ∙ Kishimoto, T. ∙ Nemoto, T. ... Two-photon excitation imaging of exocytosis and endocytosis and determination of their spatial organization Adv. Drug Deliv. Rev. 2006; 58:850-877 Crossref Scopus (37) PubMed Google Scholar
  27. [29]29. Kasai, K. ∙ Fujita, T. ∙ Gomi, H. ... Docking is not a prerequisite but a temporal constraint for fusion of secretory granules Traffic. 2008; 9:1191-1203 Crossref Scopus (93) PubMed Google Scholar
  28. [30]30. Kasai, H. ∙ Hatakeyama, H. ∙ Ohno, M. ... Exocytosis in islet beta-cells Adv. Exp. Med. Biol. 2010; 654:305-338 Crossref Scopus (32) PubMed Google Scholar
  29. [31]31. Kishimoto, T. ∙ Liu, T.T. ∙ Hatakeyama, H. ... Sequential compound exocytosis of large dense-core vesicles in PC12 cells studied with TEPIQ (two-photon extracellular polar-tracer imaging-based quantification) analysis J. Physiol. 2005; 568:905-915 Crossref Scopus (39) PubMed Google Scholar
  30. [32]32. Kishimoto, T. ∙ Kimura, R. ∙ Liu, T.-T. ... Vacuolar sequential exocytosis of large dense-core vesicles in adrenal medulla EMBO J. 2006; 25:673-682 Crossref Scopus (39) PubMed Google Scholar
  31. [33]33. Kuner, T. ∙ Li, Y. ∙ Gee, K.R. ... Photolysis of a caged peptide reveals rapid action of N-ethylmaleimide sensitive factor before neurotransmitter release Proc. Natl. Acad. Sci. USA. 2008; 105:347-352 Crossref Scopus (38) PubMed Google Scholar
  32. [34]34. Lang, J. Molecular mechanisms and regulation of insulin exocytosis as a paradigm of endocrine secretion Eur. J. Biochem. 1999; 259:3-17 Crossref Scopus (288) PubMed Google Scholar
  33. [35]35. Martin, T.F.J. Stages of regulated exocytosis Trends Cell Biol. 1997; 7:271-276

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Authors
Takahashi, N., Hatakeyama, H., Okado, H., Noguchi, J., Ohno, M., & Kasai, H.
Affiliations
  • Japan Science and Technology Agency (JP)
  • National Institute for Physiological Sciences (JP)
  • Center for Disease Biology and Integrative Medicine, The University of Tokyo (JP)
Corresponding author
  • Takahashi, Noriko
  • Kasai, Haruo
ORCID
Venue
Cell Metabolism
Publisher
Elsevier BV
Published
2010-07
Volume
12
Issue
1
Pages
19-29
ISSN
1550-4131 (ISSN-L)
DOI
10.1016/j.cmet.2010.05.013
OpenAlex
W2094356651
Cited by
70
References
3
Citations by year
2025: 2 · 2024: 4 · 2023: 1 · 2022: 3 · 2021: 3 · 2020: 2 · 2019: 3 · 2018: 1 · 2017: 7 · 2016: 3 · 2015: 8 · 2014: 6 · 2013: 5 · 2012: 10
FWCI
3.78
Type
journal article
Language
en
Open access
Licence
Locations
  • Cell Metabolism (publishedVersion)
  • PubMed (publishedVersion)
Funders
Japan Society for the Promotion of Science (21500367)
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Cell Biology › Cellular transport and secretion
  • Health Sciences › Medicine › Surgery › Pancreatic function and diabetes
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Extracellular vesicles in disease
Keywords
SNARE proteins, SNARE complex assembly, exocytosis, insulin exocytosis, pancreatic beta cells, pancreatic islets, SNAP-25, syntaxin 1A, calcium-triggered exocytosis, secretory vesicle fusion, FRET probes, two-photon imaging
MeSH terms
Animals, Calcium, Cell Membrane, Exocytosis, Insulin, Mice, Inbred ICR, Time Factors, Secretory Vesicles, Fluorescence Resonance Energy Transfer, RNA Interference, RNA, Small Interfering, Cell Line, Tumor, Insulin-Secreting Cells, SNARE Proteins, Synaptosomal-Associated Protein 25, Syntaxin 1, Mice, Rats

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