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. 2019 Jan 22;9:300. doi: 10.1038/s41598-018-36391-3

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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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Characterization of recombinant MUP20 proteins. (a) RP-HPLC analysis of recombinant wild type MUP. An aliquot (20 μg) of protein was applied to a Vydac C4 analytical column eluted with a linear acetonitrile-0.1% TFA gradient (—) at a flow rate of 0.8 ml/min. The protein material corresponding to the major peak in the chromatograms was collected and subjected to MS analysis (Supplementary Material S3). (b) Far-UV CD spectra of wild-type and mutant MUPs. CD spectra of recombinant wild type MUP (black line), and MUP mutants: L88Q (blue line), L124V (red line) and Y103R (green line). All measurements were carried out at 25 ± 0.1 °C 10 mM Na₂HPO₄ pH 7.4, 0.15 M NaCl, at a protein concentration of 0.1 mg/ml.