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. 2020 Apr 22;10:6787. doi: 10.1038/s41598-020-63926-4

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© The Author(s) 2020

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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Tks5α close neighbors belong to an integrative network related to actin cytoskeleton reorganization. MDA-MB-231 cells expressing Tks5-BirA* or BirA* were seeded on gelatin-coated plates and treated with biotin for 16 h. Biotinylated proteins were isolated by affinity capture and identified by mass spectrometry. (A) Volcano plot showing the differential level (Log2) and the p-value (-Log10), on x and y axis, respectively, for Tks5-BirA* versus BirA*. The full line is indicative of protein hits obtained at a permutation false discovery rate (pFDR) of 1%. Data result from two different experiments processed three times. Red and orange spots highlight proteins previously associated with invadosomes and proteins for which a family member has been associated with invadosomes, respectively. Green spots highlight proteins associated with cancer invasion/metastasis, outside of invadosome-associated proteins. For more details, refer to Supplementary Table S1. (B) Proteins biotinylated by Tks5-BirA*, including the validated candidate proteins (WASL and NCK1 which appear in italic) were integrated in a protein-protein interaction network based on physical and functional interactions per the STRING database. Markov Clustering algorithm has been applied. Proteins belonging to a same cluster have an identical color; the main core cluster is in red and gold. Line thickness indicates the strength/confidence of data support. Dotted lines mark the edges of the clusters. Among disconnected proteins, factors linked to actin organization are striped. The bait Tks5/SH3PXD2A is highlighted in bold. (C) Treemap representing the top biological processes represented among proteins biotinylated by Tks5-BirA*, as defined in (B), using PANTHER classification system for analysis and REVIGO for summarization and visualization. Each rectangle is a single cluster representative. The representatives are joined into ‘superclusters’ of loosely related terms, visualized with different colors. Size of the rectangles reflects the p-value of the GO term. (D) Proteins biotinylated by Tks5-BirA* (B) categorized as GO “Actin cytoskeleton organization” by PANTHER classification system.