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The impact of folding modes and deuteration on the atomic resolution structure of hen egg-white lysozyme
The biological function of a protein is intimately related to its structure and dynamics, which in turn are determined by the way in which it has been folded. In vitro refolding is commonly used for the recovery of recombinant proteins that are expressed in the form of inclusion bodies and is of cen...
Autores principales: | , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
International Union of Crystallography
2021
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8647175/ https://www.ncbi.nlm.nih.gov/pubmed/34866613 http://dx.doi.org/10.1107/S2059798321010950 |
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author | Ramos, Joao Laux, Valerie Haertlein, Michael Forsyth, V. Trevor Mossou, Estelle Larsen, Sine Langkilde, Annette E. |
author_facet | Ramos, Joao Laux, Valerie Haertlein, Michael Forsyth, V. Trevor Mossou, Estelle Larsen, Sine Langkilde, Annette E. |
author_sort | Ramos, Joao |
collection | PubMed |
description | The biological function of a protein is intimately related to its structure and dynamics, which in turn are determined by the way in which it has been folded. In vitro refolding is commonly used for the recovery of recombinant proteins that are expressed in the form of inclusion bodies and is of central interest in terms of the folding pathways that occur in vivo. Here, biophysical data are reported for in vitro-refolded hydrogenated hen egg-white lysozyme, in combination with atomic resolution X-ray diffraction analyses, which allowed detailed comparisons with native hydrogenated and refolded perdeuterated lysozyme. Distinct folding modes are observed for the hydrogenated and perdeuterated refolded variants, which are determined by conformational changes to the backbone structure of the Lys97–Gly104 flexible loop. Surprisingly, the structure of the refolded perdeuterated protein is closer to that of native lysozyme than that of the refolded hydrogenated protein. These structural differences suggest that the observed decreases in thermal stability and enzymatic activity in the refolded perdeuterated and hydrogenated proteins are consequences of the macromolecular deuteration effect and of distinct folding dynamics, respectively. These results are discussed in the context of both in vitro and in vivo folding, as well as of lysozyme amyloidogenesis. |
format | Online Article Text |
id | pubmed-8647175 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2021 |
publisher | International Union of Crystallography |
record_format | MEDLINE/PubMed |
spelling | pubmed-86471752021-12-16 The impact of folding modes and deuteration on the atomic resolution structure of hen egg-white lysozyme Ramos, Joao Laux, Valerie Haertlein, Michael Forsyth, V. Trevor Mossou, Estelle Larsen, Sine Langkilde, Annette E. Acta Crystallogr D Struct Biol Research Papers The biological function of a protein is intimately related to its structure and dynamics, which in turn are determined by the way in which it has been folded. In vitro refolding is commonly used for the recovery of recombinant proteins that are expressed in the form of inclusion bodies and is of central interest in terms of the folding pathways that occur in vivo. Here, biophysical data are reported for in vitro-refolded hydrogenated hen egg-white lysozyme, in combination with atomic resolution X-ray diffraction analyses, which allowed detailed comparisons with native hydrogenated and refolded perdeuterated lysozyme. Distinct folding modes are observed for the hydrogenated and perdeuterated refolded variants, which are determined by conformational changes to the backbone structure of the Lys97–Gly104 flexible loop. Surprisingly, the structure of the refolded perdeuterated protein is closer to that of native lysozyme than that of the refolded hydrogenated protein. These structural differences suggest that the observed decreases in thermal stability and enzymatic activity in the refolded perdeuterated and hydrogenated proteins are consequences of the macromolecular deuteration effect and of distinct folding dynamics, respectively. These results are discussed in the context of both in vitro and in vivo folding, as well as of lysozyme amyloidogenesis. International Union of Crystallography 2021-11-17 /pmc/articles/PMC8647175/ /pubmed/34866613 http://dx.doi.org/10.1107/S2059798321010950 Text en © Joao Ramos et al. 2021 https://creativecommons.org/licenses/by/4.0/This is an open-access article distributed under the terms of the Creative Commons Attribution (CC-BY) Licence, which permits unrestricted use, distribution, and reproduction in any medium, provided the original authors and source are cited. |
spellingShingle | Research Papers Ramos, Joao Laux, Valerie Haertlein, Michael Forsyth, V. Trevor Mossou, Estelle Larsen, Sine Langkilde, Annette E. The impact of folding modes and deuteration on the atomic resolution structure of hen egg-white lysozyme |
title | The impact of folding modes and deuteration on the atomic resolution structure of hen egg-white lysozyme |
title_full | The impact of folding modes and deuteration on the atomic resolution structure of hen egg-white lysozyme |
title_fullStr | The impact of folding modes and deuteration on the atomic resolution structure of hen egg-white lysozyme |
title_full_unstemmed | The impact of folding modes and deuteration on the atomic resolution structure of hen egg-white lysozyme |
title_short | The impact of folding modes and deuteration on the atomic resolution structure of hen egg-white lysozyme |
title_sort | impact of folding modes and deuteration on the atomic resolution structure of hen egg-white lysozyme |
topic | Research Papers |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8647175/ https://www.ncbi.nlm.nih.gov/pubmed/34866613 http://dx.doi.org/10.1107/S2059798321010950 |
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