Thermostabilization of VPR, a kinetically stable cold adapted subtilase, via multiple proline substitutions into surface loops
| dc.contributor | Háskóli Íslands | en_US |
| dc.contributor | University of Iceland | en_US |
| dc.contributor.author | Óskarsson, Kristinn Ragnar | |
| dc.contributor.author | Sævarsson, Arnór Freyr | |
| dc.contributor.author | Kristjánsson, Magnús M. | |
| dc.contributor.department | Raunvísindastofnun (HÍ) | en_US |
| dc.contributor.department | Science Institute (UI) | en_US |
| dc.contributor.school | Verkfræði- og náttúruvísindasvið (HÍ) | en_US |
| dc.contributor.school | School of Engineering and Natural Sciences (UI) | en_US |
| dc.date.accessioned | 2020-11-02T14:47:48Z | |
| dc.date.available | 2020-11-02T14:47:48Z | |
| dc.date.issued | 2020-01-23 | |
| dc.description | Publisher's version (útgefin grein) | en_US |
| dc.description.abstract | Protein stability is a widely studied topic, there are still aspects however that need addressing. In this paper we examined the effects of multiple proline substitutions into loop regions of the kinetically stable proteinase K-like serine protease VPR, using the thermostable structural homologue AQUI as a template. Four locations for proline substitutions were chosen to imitate the structure of AQUI. Variants were produced and characterized using differential scanning calorimetry (DSC), circular dichroism (CD), steady state fluorescence, acrylamide fluorescence quenching and thermal inactivation experiments. The final product VPRΔC_N3P/I5P/N238P/T265P was greatly stabilized which was achieved without any noticeable detrimental effects to the catalytic efficiency of the enzyme. This stabilization seems to be derived from the conformation restrictive properties of the proline residue in its ability to act as an anchor point and strengthen pre-existing interactions within the protein and allowing for these interactions to prevail when thermal energy is applied to the system. In addition, the results underline the importance of the synergy between distant local protein motions needed to result in stabilizing effects and thus giving an insight into the nature of the stability of VPR, its unfolding landscape and how proline residues can infer kinetic stability onto protein structures. | en_US |
| dc.description.sponsorship | This work was supported by The Icelandic Research Fund [Grant Number 162977-051]. | en_US |
| dc.description.version | Peer Reviewed | en_US |
| dc.format.extent | 1045 | en_US |
| dc.identifier.citation | Óskarsson, K.R., Sævarsson, A.F. & Kristjánsson, M.M. Thermostabilization of VPR, a kinetically stable cold adapted subtilase, via multiple proline substitutions into surface loops. Sci Rep 10, 1045 (2020). https://doi.org/10.1038/s41598-020-57873-3 | en_US |
| dc.identifier.doi | 10.1038/s41598-020-57873-3 | |
| dc.identifier.issn | 2045-2322 | |
| dc.identifier.journal | Scientific Reports | en_US |
| dc.identifier.uri | https://hdl.handle.net/20.500.11815/2157 | |
| dc.language.iso | en | en_US |
| dc.publisher | Springer Science and Business Media LLC | en_US |
| dc.relation.ispartofseries | Scientific Reports;10(1) | |
| dc.relation.url | https://www.nature.com/articles/s41598-020-57873-3 | en_US |
| dc.rights | info:eu-repo/semantics/openAccess | en_US |
| dc.subject | Protein stability | en_US |
| dc.subject | VPR | en_US |
| dc.subject | AQUI | en_US |
| dc.subject | Prótín | en_US |
| dc.subject | Lífefnafræði | en_US |
| dc.title | Thermostabilization of VPR, a kinetically stable cold adapted subtilase, via multiple proline substitutions into surface loops | en_US |
| dc.type | info:eu-repo/semantics/article | en_US |
| dcterms.license | 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/. | en_US |
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