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. 2019 Nov 27;9:17662. doi: 10.1038/s41598-019-54198-8

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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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CD spectra of WT NhaA and mutant R245A-R250A. The respective proteins (4 µM) were affinity-purified and re-suspended in dialysis buffer titrated to pH 7.5 with Tris base. The CD spectra were recorded on a JASCO J-810 CD Spectro-polarimeter (JASCO, Inc., Japan), using the supplied Spectra Manager software. (a) Comparison of CD spectra for the WT and mutant R245A-R250A at 4 °C. (b) Unfolding transition midpoint (T_m) values were determined from the corresponding inflection points. Traces are representative from three different measurements.