A multiscale in situ time-resolved study of the nano- to millisecond structural dynamics during protein crystallization
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| Název: | A multiscale in situ time-resolved study of the nano- to millisecond structural dynamics during protein crystallization |
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| Autoři: | Beck, Christian, Mosca, Ilaria, Miñarro, Laura M., Sohmen, Benedikt, Buchholz, Cara, Maier, Ralph, Reichart, Lara Franziska, Grundel, Anna Carlotta, Bäuerle, Famke, Nasro, Roody, Banks, Hadra, Christmann, Simon, Pastryk, Kai-Florian, Farago, Bela, Czakkel, Orsolya, Prévost, Sylvain, Gerlach, Alexander, Grimaldo, Marco, Roosen-Runge, Felix, Matsarskaia, Olga, Schreiber, Frank, Seydel, Tilo |
| Přispěvatelé: | Lund University, Faculty of Science, Department of Chemistry, Physical and theoretical chemistry, Physical Chemistry, Lunds universitet, Naturvetenskapliga fakulteten, Kemiska institutionen, Enheten för fysikalisk och teoretisk kemi, Fysikalisk kemi, Originator |
| Zdroj: | Journal of Applied Crystallography. 58(Pt 3):845-858 |
| Témata: | Natural Sciences, Chemical Sciences, Physical Chemistry (including Surface- and Colloid Chemistry), Naturvetenskap, Kemi, Fysikalisk kemi (Här ingår: Yt- och kolloidkemi) |
| Popis: | 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 toreveal 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. |
| Přístupová URL adresa: | https://doi.org/10.1107/S160057672500353X |
| Databáze: | SwePub |
Nájsť tento článok vo Web of Science | Abstrakt: | 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 toreveal 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. |
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| ISSN: | 00218898 16005767 |
| DOI: | 10.1107/S160057672500353X |