Article Dans Une Revue Journal of Applied Crystallography Année : 2025

A multiscale in situ time-resolved study of the nano- to millisecond structural dynamics during protein crystallization

Christian Beck
Ilaria Mosca
Laura Miñarro
  • Fonction : Auteur
Benedikt Sohmen
  • Fonction : Auteur
Cara Buchholz
  • Fonction : Auteur
Ralph Maier
  • Fonction : Auteur
Lara Franziska Reichart
  • Fonction : Auteur
Anna Carlotta Grundel
  • Fonction : Auteur
Famke Bäuerle
Roody Nasro
  • Fonction : Auteur
Hadra Banks
Simon Christmann
  • Fonction : Auteur
Kai-Florian Pastryk
  • Fonction : Auteur
Bela Farago
  • Fonction : Auteur
Orsolya Czakkel
  • Fonction : Auteur
Sylvain Prévost
  • Fonction : Auteur
Alexander Gerlach
Marco Grimaldo
  • Fonction : Auteur
Felix Roosen-Runge
  • Fonction : Auteur
Olga Matsarskaia
  • Fonction : Auteur
Frank Schreiber
Tilo Seydel

Résumé

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 LaCl 3 . 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 3 model system with the crystallization behavior of β-lactoglobulin–CdCl 2 , which includes a metastable intermediate state.

Fichier principal
Vignette du fichier
JApplCryst_vol58_p845_2025_jo5120_open_access_CC_BY.pdf (4.25 Mo) Télécharger le fichier
JApplCryst_vol58_p845_2025_jo5120_Supplementary_Material.pdf (1.73 Mo) Télécharger le fichier
Origine Fichiers éditeurs autorisés sur une archive ouverte
Licence

Dates et versions

hal-05091120 , version 1 (03-06-2025)

Licence

Identifiants

Citer

Christian Beck, Ilaria Mosca, Laura Miñarro, Benedikt Sohmen, Cara Buchholz, et al.. A multiscale in situ time-resolved study of the nano- to millisecond structural dynamics during protein crystallization. Journal of Applied Crystallography, 2025, 58 (3), ⟨10.1107/S160057672500353X⟩. ⟨hal-05091120⟩

Collections

52 Consultations
102 Téléchargements

Altmetric

Partager

  • More