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. 2020 Aug 6;10:13297. doi: 10.1038/s41598-020-69990-0

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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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Modification of mtDod-mmACP, mtDod-mmACP-H8 and mmACP by Sfp with fluorescent CoA. L: ladder. NC: negative control reaction without Sfp. Eq.: molar equivalents of mmACP loaded onto the SDS-PAGE gel. (a) Top: Coomassie stained SDS-PAGE gel of the reaction solution and negative controls. Sfp, runs at an apparent molecular weight of slightly above 25 kDa, mtDod-mmACP-H8 slightly below 25 kDa and mmACP slightly below 15 kDa. Unmodified mmACP and mtDod-mmACP-H8 (negative controls) show lower apparent molecular weights indicating a different running behaviour between CoA-488 modified and unmodified proteins. Bottom: In-gel fluorescence taken before Coomassie staining. Only proteins modified with the CoA-488 are visible. Higher bands represent labelled mtDod-mmACP-H8 and lower bands labelled mmACP. (b) Top: Coomassie stained SDS-PAGE gel of the reaction solutions of mtDod-mmACP-H8 and mtDod-mmACP after 1 h and 2 h. Bottom: in-gel fluorescence taken before Coomassie staining. For both constructs, fluorescence intensity increases at longer reaction times. For uncropped images and mtDod-mmACP blots see Supplementary Fig. S9.