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. 2018 Apr 12;8:5903. doi: 10.1038/s41598-018-24068-w

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© The Author(s) 2018

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 Aco1 C-terminal and N-terminal domains interact in vivo. (A) Schematic representation of aconitase GST and HA fusion proteins. N terminal domains, green; C terminal domain, light blue; MTS, purple; GST tag, orange; HA tag, red. (B) Aco1 C and N terminal fusions are expressed in yeast. Total cell lysates of yeast cultures expressing the indicated GST and HA fusions were subjected to Western blot analysis using either anti–GST or anti–HA antiserum. Arrows indicate bands of GST-C or GST-C∆6 fusions (“a”), HA-Aco1 fusions (“b”) and HA-N fusions (“c”). (C) GST-Aco1-C (C terminal domain) can pull-down full length aconitase and the N-terminal domains. Lysates of yeast cells expressing the indicated fusion proteins were incubated with glutathione beads which were subsequently washed and resuspended in SDS containing loading buffer. Samples were subjected to Western blot analysis using the indicated antisera. Arrows indicate bands of GST-C or GST-C∆6 fusions (“a”), HA-Aco1 fusions (“b”) and HA-N fusions (“c”).