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. 2017 Aug 18;7:8700. doi: 10.1038/s41598-017-09162-9

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© The Author(s) 2017

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Multiple GTPase cycles are not required to maintain membrane tethering. (A) Single-vesicle images upon donor and acceptor excitation before and after removing GTP and Mg²⁺. Three independent experiments all indicated that the tethered vesicles did not dissociate. (B) Representative time traces observed in a buffer-exchange experiment. Buffer lacking GTP and Mg²⁺ was injected into the detection chamber during incubation of donor vesicles in the presence of GTP and Mg²⁺. Experiments were performed using proteoliposomes with a protein-to-lipid ratio of 1:200.