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Quantitative GTPase affinity purification identifies Rho family protein interaction partners

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Item Type:Article
Title:Quantitative GTPase affinity purification identifies Rho family protein interaction partners
Creators Name:Paul, F. and Zauber, H. and von Berg, L. and Rocks, O. and Daumke, O. and Selbach, M.
Abstract:Although Rho GTPases are essential molecular switches involved in many cellular processes, an unbiased experimental comparison of their interaction partners was not yet performed. Here, we develop quantitative GTPase affinity purification (qGAP) to systematically identify interaction partners of six Rho GTPases (Cdc42, Rac1, RhoA, RhoB, RhoC, RhoD) depending on their nucleotide loading state. The method works with cell line or tissue-derived protein lysates in combination with SILAC-based or label free quantification, respectively. We demonstrate that qGAP identifies known and novel binding partners that can be validated in an independent assay. Our interaction network for six Rho GTPases contains many novel binding partners, reveals highly promiscuous interaction of several effectors and mirrors evolutionary relationships among Rho GTPases.
Keywords:Rho GTPase, Proteomics, Signaling, Protein-Protein Interaction, Animals, Mice
Source:Molecular & Cellular Proteomics
ISSN:1535-9476
Publisher:American Society for Biochemistry and Molecular Biology (U.S.A.)
Volume:16
Number:1
Page Range:73-85
Date:1 January 2017
Official Publication:https://doi.org/10.1074/mcp.M116.061531
PubMed:View item in PubMed

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