Close is not enough: SNARE-dependent membrane fusion requires an active mechanism that transduces force to membrane anchors Journal Article
| Authors: | Mcnew, J. A.; Weber, T.; Parlati, F.; Johnston, R. J.; Melia, T. J.; Söllner, T. H.; Rothman, J. E. |
| Article Title: | Close is not enough: SNARE-dependent membrane fusion requires an active mechanism that transduces force to membrane anchors |
| Abstract: | Is membrane fusion an essentially passive or an active process? It could be that fusion proteins simply need to pin two bilayers together long enough, and the bilayers could do the rest spontaneously. Or, it could be that the fusion proteins play an active role after pinning two bilayers, exerting force in the bilayer in one or another way to direct the fusion process. To distinguish these alternatives, we replaced one or both of the peptidic membrane anchors of exocytic vesicle (v)- and target membrane (t)-SNAREs (soluble N-ethylmaleimide-sensitive fusion protein [NSF] attachment protein [SNAP] receptor) with covalently attached lipids. Replacing either anchor with a phospholipid prevented fusion of liposomes by the isolated SNAREs, but still allowed assembly of trans-SNARE complexes docking vesicles. This result implies an active mechanism; if fusion occurred passively, simply holding the bilayers together long enough would have been sufficient. Studies using polyisoprenoid anchors ranging from 15-55 carbons and multiple phospholipid- containing anchors reveal distinct requirements for anchors of v- and t- SNAREs to function: v-SNAREs require anchors capable of spanning both leaflets, whereas t-SNAREs do not, so long as the anchor is sufficiently hydrophobic. These data, together with previous results showing fusion is inhibited as the length of the linker connecting the helical bundle- containing rod of the SNARE complex to the anchors is increased (McNew, J.A., T. Weber, D.M. Engelman, T.H. Sollner, and J.E. Rothman. 1999. Mol. Cell. 4:415-421), suggests a model in which one activity of the SNARE complex promoting fusion is to exert force on the anchors by pulling on the linkers. This motion would lead to the simultaneous inward movement of lipids from both bilayers, and in the case of the v-SNARE, from both leaflets. |
| Keywords: | nerve tissue proteins; membrane proteins; cell membrane; cytoplasm; protein structure, tertiary; protein structure; structure analysis; models, chemical; liposome; liposomes; lipid bilayers; lipid bilayer; antigens, surface; phospholipids; membrane vesicle; vesicular transport proteins; cross-linking reagents; vesicular transport; membrane fusion; covalent bond; snare protein; syntaxin 1; r-snare proteins; snare proteins; terpenes; synaptosomal-associated protein 25; glycosylphosphatidylinositols; priority journal; article; isoprene; lipid anchor; lipid mixing |
| Journal Title: | Journal of Cell Biology |
| Volume: | 150 |
| Issue: | 1 |
| ISSN: | 0021-9525 |
| Publisher: | Rockefeller University Press |
| Date Published: | 2000-07-10 |
| Start Page: | 105 |
| End Page: | 117 |
| Language: | English |
| DOI: | 10.1083/jcb.150.1.105 |
| PUBMED: | 10893260 |
| PROVIDER: | scopus |
| PMCID: | PMC2185554 |
| DOI/URL: | |
| Notes: | Export Date: 18 November 2015 -- Source: Scopus |
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