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. 2018 Jul 4;8:10124. doi: 10.1038/s41598-018-28290-4

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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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CarD undergoes reversible thermal folding in solution. (a) CD spectra obtained during thermal denaturation of CarD (10 μM). CarD was heated from 20 to 95 °C and then cooled from 95 to 20 °C. For clarity, only the representative CD spectra acquired at 20 °C, 55 °C and 95 °C during both heating and cooling (labeled with ‘_R’) have been plotted. The CD spectra reveal partial unfolding at elevated temperatures and refolding upon cooling with the presence of isobestic/isodichroic point at 205 nm. (b) The plot showing variations in secondary structural content of CarD with change in temperature. With the increase in temperature, the percentage of α-helices decreases and the percentage of anti-parallel β-sheets increases significantly.