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. 2002 Nov;83(5):2522–2527. doi: 10.1016/s0006-3495(02)75263-6

SNAREs in opposing bilayers interact in a circular array to form conducting pores.

Sang-Joon Cho 1, Marie Kelly 1, Katherine T Rognlien 1, Jin Ah Cho 1, J K Heinrich Hörber 1, Bhanu P Jena 1
PMCID: PMC1302338  PMID: 12414686

Abstract

The process of fusion at the nerve terminal is mediated via a specialized set of proteins in the synaptic vesicles and the presynaptic membrane. Three soluble N-ethylmaleimide-sensitive factor (NSF)-attachment protein receptors (SNAREs) have been implicated in membrane fusion. The structure and arrangement of these SNAREs associated with lipid bilayers were examined using atomic force microscopy. A bilayer electrophysiological setup allowed for measurements of membrane conductance and capacitance. Here we demonstrate that the interaction of these proteins to form a fusion pore is dependent on the presence of t-SNAREs and v-SNARE in opposing bilayers. Addition of purified recombinant v-SNARE to a t-SNARE-reconstituted lipid membrane increased only the size of the globular t-SNARE oligomer without influencing the electrical properties of the membrane. However when t-SNARE vesicles were added to a v-SNARE membrane, SNAREs assembles in a ring pattern and a stepwise increase in capacitance, and increase in conductance were observed. Thus, t- and v-SNAREs are required to reside in opposing bilayers to allow appropriate t-/v-SNARE interactions leading to membrane fusion.

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Selected References

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  1. Chan D. C., Kim P. S. HIV entry and its inhibition. Cell. 1998 May 29;93(5):681–684. doi: 10.1016/s0092-8674(00)81430-0. [DOI] [PubMed] [Google Scholar]
  2. Cho Sang-Joon, Jeftinija Ksenija, Glavaski Aleksandra, Jeftinija Srdija, Jena Bhanu P., Anderson Lloyd L. Structure and dynamics of the fusion pores in live GH-secreting cells revealed using atomic force microscopy. Endocrinology. 2002 Mar;143(3):1144–1148. doi: 10.1210/endo.143.3.8773. [DOI] [PubMed] [Google Scholar]
  3. Cho Sang-Joon, Quinn Anthony S., Stromer Marvin H., Dash Sudhansu, Cho Jinah, Taatjes Douglas J., Jena Bhanu P. Structure and dynamics of the fusion pore in live cells. Cell Biol Int. 2002;26(1):35–42. doi: 10.1006/cbir.2001.0849. [DOI] [PubMed] [Google Scholar]
  4. Cho Sang-Joon, Sattar A. K. M. Abdus, Jeong Eun-Hwan, Satchi Mylan, Cho Jin Ah, Dash Sudhansu, Mayes Mary Sue, Stromer Marvin H., Jena Bhanu P. Aquaporin 1 regulates GTP-induced rapid gating of water in secretory vesicles. Proc Natl Acad Sci U S A. 2002 Mar 26;99(7):4720–4724. doi: 10.1073/pnas.072083499. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Cohen F. S., Niles W. D. Reconstituting channels into planar membranes: a conceptual framework and methods for fusing vesicles to planar bilayer phospholipid membranes. Methods Enzymol. 1993;220:50–68. doi: 10.1016/0076-6879(93)20073-c. [DOI] [PubMed] [Google Scholar]
  6. Fasshauer D., Otto H., Eliason W. K., Jahn R., Brünger A. T. Structural changes are associated with soluble N-ethylmaleimide-sensitive fusion protein attachment protein receptor complex formation. J Biol Chem. 1997 Oct 31;272(44):28036–28041. doi: 10.1074/jbc.272.44.28036. [DOI] [PubMed] [Google Scholar]
  7. Hanson P. I., Roth R., Morisaki H., Jahn R., Heuser J. E. Structure and conformational changes in NSF and its membrane receptor complexes visualized by quick-freeze/deep-etch electron microscopy. Cell. 1997 Aug 8;90(3):523–535. doi: 10.1016/s0092-8674(00)80512-7. [DOI] [PubMed] [Google Scholar]
  8. Hayashi T., Yamasaki S., Nauenburg S., Binz T., Niemann H. Disassembly of the reconstituted synaptic vesicle membrane fusion complex in vitro. EMBO J. 1995 May 15;14(10):2317–2325. doi: 10.1002/j.1460-2075.1995.tb07226.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Jeong E. H., Webster P., Khuong C. Q., Abdus Sattar A. K., Satchi M., Jena B. P. The native membrane fusion machinery in cells. Cell Biol Int. 1998;22(9-10):657–670. doi: 10.1006/cbir.1999.0394. [DOI] [PubMed] [Google Scholar]
  10. Kelly M. L., Woodbury D. J. Ion channels from synaptic vesicle membrane fragments reconstituted into lipid bilayers. Biophys J. 1996 Jun;70(6):2593–2599. doi: 10.1016/S0006-3495(96)79830-2. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Rothman J. E. Mechanisms of intracellular protein transport. Nature. 1994 Nov 3;372(6501):55–63. doi: 10.1038/372055a0. [DOI] [PubMed] [Google Scholar]
  12. Schneider S. W., Sritharan K. C., Geibel J. P., Oberleithner H., Jena B. P. Surface dynamics in living acinar cells imaged by atomic force microscopy: identification of plasma membrane structures involved in exocytosis. Proc Natl Acad Sci U S A. 1997 Jan 7;94(1):316–321. doi: 10.1073/pnas.94.1.316. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Sutton R. B., Fasshauer D., Jahn R., Brunger A. T. Crystal structure of a SNARE complex involved in synaptic exocytosis at 2.4 A resolution. Nature. 1998 Sep 24;395(6700):347–353. doi: 10.1038/26412. [DOI] [PubMed] [Google Scholar]
  14. Söllner T., Bennett M. K., Whiteheart S. W., Scheller R. H., Rothman J. E. A protein assembly-disassembly pathway in vitro that may correspond to sequential steps of synaptic vesicle docking, activation, and fusion. Cell. 1993 Nov 5;75(3):409–418. doi: 10.1016/0092-8674(93)90376-2. [DOI] [PubMed] [Google Scholar]
  15. Söllner T., Whiteheart S. W., Brunner M., Erdjument-Bromage H., Geromanos S., Tempst P., Rothman J. E. SNAP receptors implicated in vesicle targeting and fusion. Nature. 1993 Mar 25;362(6418):318–324. doi: 10.1038/362318a0. [DOI] [PubMed] [Google Scholar]
  16. Weber T., Zemelman B. V., McNew J. A., Westermann B., Gmachl M., Parlati F., Söllner T. H., Rothman J. E. SNAREpins: minimal machinery for membrane fusion. Cell. 1998 Mar 20;92(6):759–772. doi: 10.1016/s0092-8674(00)81404-x. [DOI] [PubMed] [Google Scholar]
  17. Woodbury D. J., Miller C. Nystatin-induced liposome fusion. A versatile approach to ion channel reconstitution into planar bilayers. Biophys J. 1990 Oct;58(4):833–839. doi: 10.1016/S0006-3495(90)82429-2. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Woodbury D. J. Nystatin/ergosterol method for reconstituting ion channels into planar lipid bilayers. Methods Enzymol. 1999;294:319–339. doi: 10.1016/s0076-6879(99)94020-x. [DOI] [PubMed] [Google Scholar]

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