A multiscale in situ time-resolved study of the nano- to millisecond structural dynamics during protein crystallization
(2025) In Journal of Applied Crystallography 58(Pt 3). p.845-858- Abstract
- Protein crystallization is key to determining the structure of proteins at atomic resolution. It can occur naturally, including in pathological pathways, for instance with aquaporin and γ-crystallin proteins. A fundamental understanding of the underlying crystallization process is both technologically and biologically relevant. A multitechnique approach is employed here to investigate protein crystallization in situ, allowing us to assess the evolution of the liquid suspension and crystallite structure as well as protein diffusion during the crystallization process. The wide range of methods probe the sample on ångström to millimetre length scales, accessing nanosecond to millisecond dynamics information while acquiring data with... (More)
- Protein crystallization is key to determining the structure of proteins at atomic resolution. It can occur naturally, including in pathological pathways, for instance with aquaporin and γ-crystallin proteins. A fundamental understanding of the underlying crystallization process is both technologically and biologically relevant. A multitechnique approach is employed here to investigate protein crystallization in situ, allowing us to assess the evolution of the liquid suspension and crystallite structure as well as protein diffusion during the crystallization process. The wide range of methods probe the sample on ångström to millimetre length scales, accessing nanosecond to millisecond dynamics information while acquiring data with minute-timescale kinetic resolution during crystallization. This process takes several hours from an initial state of monomers or small clusters until the presence of large crystallites. Employing neutron spectroscopy allows us to distinguish different crystallization pathways and to reveal the presence of coexisting clusters during the entire crystallization process. We demonstrate the multitechnique approach on human serum albumin (HSA) proteins crystallized from aqueous solution in the presence of LaCl3. For this system, the crystallization kinetics can be consistently described by a sigmoid function across all methods, and the kinetics can be controlled by the salt concentration. Moreover, we compare the HSA–LaCl3 model system with the crystallization behavior of β-lactoglobulin–CdCl2, which includes a metastable intermediate state. (Less)
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https://lup.lub.lu.se/record/d0ae3572-2fc2-4f01-ad41-8e740ff5690c
- author
- organization
- publishing date
- 2025-06-01
- type
- Contribution to journal
- publication status
- published
- subject
- keywords
- diffusion, in situ protein crystallization, neutron spectroscopy, small-angle neutron scattering, structural dynamics
- in
- Journal of Applied Crystallography
- volume
- 58
- issue
- Pt 3
- pages
- 14 pages
- publisher
- International Union of Crystallography
- external identifiers
-
- scopus:105007508725
- pmid:40475925
- ISSN
- 0021-8898
- DOI
- 10.1107/S160057672500353X
- language
- English
- LU publication?
- yes
- additional info
- Publisher Copyright: © 2025 International Union of Crystallography. All rights reserved.
- id
- d0ae3572-2fc2-4f01-ad41-8e740ff5690c
- date added to LUP
- 2025-08-12 23:17:06
- date last changed
- 2025-08-21 09:57:40
@article{d0ae3572-2fc2-4f01-ad41-8e740ff5690c,
abstract = {{Protein crystallization is key to determining the structure of proteins at atomic resolution. It can occur naturally, including in pathological pathways, for instance with aquaporin and γ-crystallin proteins. A fundamental understanding of the underlying crystallization process is both technologically and biologically relevant. A multitechnique approach is employed here to investigate protein crystallization <i>in situ</i>, allowing us to assess the evolution of the liquid suspension and crystallite structure as well as protein diffusion during the crystallization process. The wide range of methods probe the sample on ångström to millimetre length scales, accessing nanosecond to millisecond dynamics information while acquiring data with minute-timescale kinetic resolution during crystallization. This process takes several hours from an initial state of monomers or small clusters until the presence of large crystallites. Employing neutron spectroscopy allows us to distinguish different crystallization pathways and to reveal the presence of coexisting clusters during the entire crystallization process. We demonstrate the multitechnique approach on human serum albumin (HSA) proteins crystallized from aqueous solution in the presence of LaCl<sub>3</sub>. For this system, the crystallization kinetics can be consistently described by a sigmoid function across all methods, and the kinetics can be controlled by the salt concentration. Moreover, we compare the HSA–LaCl<sub>3</sub> model system with the crystallization behavior of β-lactoglobulin–CdCl<sub>2</sub>, which includes a metastable intermediate state.}},
author = {{Beck, Christian and Mosca, Ilaria and Miñarro, Laura M. and Sohmen, Benedikt and Buchholz, Cara and Maier, Ralph and Reichart, Lara Franziska and Grundel, Anna Carlotta and Bäuerle, Famke and Nasro, Roody and Banks, Hadra and Christmann, Simon and Pastryk, Kai-Florian and Farago, Bela and Czakkel, Orsolya and Prévost, Sylvain and Gerlach, Alexander and Grimaldo, Marco and Roosen-Runge, Felix and Matsarskaia, Olga and Schreiber, Frank and Seydel, Tilo}},
issn = {{0021-8898}},
keywords = {{diffusion; in situ protein crystallization; neutron spectroscopy; small-angle neutron scattering; structural dynamics}},
language = {{eng}},
month = {{06}},
number = {{Pt 3}},
pages = {{845--858}},
publisher = {{International Union of Crystallography}},
series = {{Journal of Applied Crystallography}},
title = {{A multiscale <i>in situ </i>time-resolved study of the nano- to millisecond structural dynamics during protein crystallization}},
url = {{http://dx.doi.org/10.1107/S160057672500353X}},
doi = {{10.1107/S160057672500353X}},
volume = {{58}},
year = {{2025}},
}