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. 2019 Jun 19;9:8725. doi: 10.1038/s41598-019-44259-3

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© The Author(s) 2019

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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The sizes of VIPP1-containing particles increase in the presence of PC:PI4P- liposomes. (a) Separation of 2 µg each of recombinant VIPP1, tVIPP1 and CGE1 on a 12% SDS-polyacrylamide gel, stained with Coomassie brilliant blue. (b) Protein–lipid overlay assay. Membranes spotted with different lipid species were incubated with the recombinant proteins followed by immunodetection of the bound proteins. Lipid species spotted were GT – glyceryl tripalmitate, DAG – diacylglycerol, PA – phosphatidic acid, PS – phosphatidylserine, PE – phosphatidylethanolamine, PC – phosphatidylcholine, PG – phosphatidylglycerol, CL – cardiolipin, PI – phosphatidylinositol, PIP – phosphatidylinositol phosphate, SM – sphingomyelin. Blue blank (Xylene Cyanol FF) is the negative control. (c) Fitted fluorescence anisotropy decay curves for VIPP1 without liposomes (none) and mixtures of VIPP1 with liposomes composed of PC, PC:PG (95:5), and PC:PI4P (95:5). (d) Best-fit values and 68% confidence intervals for the slow correlation times, θ_slow, which reflect the rotational mobility of VIPP1. Note that in most cases error bars are too small to be seen. (e) Best-fit values and 68% confidence intervals for θ_slow for the measurement shown in d and a measurement with tVIPP1. (f) Best-fit values and 68% confidence intervals for θ_slow for the measurement shown in d and a subsequent measurement performed in the presence of 10 mM Mg²⁺.