The impact of folding modes and deuteration on the atomic resolution structure of hen egg-white lysozyme
(2021) In Acta Crystallographica Section D: Structural Biology 77. p.1579-1590- Abstract
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... (More)
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.
(Less)
- author
- Ramos, Joao ; Laux, Valerie ; Haertlein, Michael ; Forsyth, V. Trevor LU ; Mossou, Estelle ; Larsen, Sine LU and Langkilde, Annette E.
- organization
- publishing date
- 2021-12
- type
- Contribution to journal
- publication status
- published
- subject
- keywords
- Deuteration, Enzymatic activity, Folding dynamics, Folding modes, Hen egg-white lysozyme, In vitro refolding, Isotope effect, Thermal stability, X-ray crystallography
- in
- Acta Crystallographica Section D: Structural Biology
- volume
- 77
- pages
- 12 pages
- publisher
- John Wiley & Sons Inc.
- external identifiers
-
- scopus:85121443981
- pmid:34866613
- ISSN
- 2059-7983
- DOI
- 10.1107/S2059798321010950
- language
- English
- LU publication?
- yes
- id
- 36e35d78-dfd3-43e0-b190-949bca30f9d7
- date added to LUP
- 2022-01-26 14:39:46
- date last changed
- 2024-06-17 03:12:28
@article{36e35d78-dfd3-43e0-b190-949bca30f9d7,
abstract = {{<p>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.</p>}},
author = {{Ramos, Joao and Laux, Valerie and Haertlein, Michael and Forsyth, V. Trevor and Mossou, Estelle and Larsen, Sine and Langkilde, Annette E.}},
issn = {{2059-7983}},
keywords = {{Deuteration; Enzymatic activity; Folding dynamics; Folding modes; Hen egg-white lysozyme; In vitro refolding; Isotope effect; Thermal stability; X-ray crystallography}},
language = {{eng}},
pages = {{1579--1590}},
publisher = {{John Wiley & Sons Inc.}},
series = {{Acta Crystallographica Section D: Structural Biology}},
title = {{The impact of folding modes and deuteration on the atomic resolution structure of hen egg-white lysozyme}},
url = {{http://dx.doi.org/10.1107/S2059798321010950}},
doi = {{10.1107/S2059798321010950}},
volume = {{77}},
year = {{2021}},
}