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
Institute of Applied Physics University of Tübingen 72076Tübingen Germany.
Institut Max von Laue-Paul Langevin, 38042Grenoble Cedex 9, France.
J Appl Crystallogr. 2025 May 29;58(Pt 3):845-858. doi: 10.1107/S160057672500353X. eCollection 2025 Jun 1.
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 , 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. 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 model system with the crystallization behavior of β-lactoglobulin-CdCl, which includes a metastable intermediate state.
蛋白质结晶是在原子分辨率下确定蛋白质结构的关键。它可以自然发生，包括在病理途径中，例如水通道蛋白和γ-晶状体蛋白。对潜在结晶过程的基本理解在技术和生物学上都具有相关性。本文采用多种技术方法来研究蛋白质结晶，使我们能够评估液体悬浮液和微晶结构的演变以及结晶过程中蛋白质的扩散。这些广泛的方法在从埃到毫米的长度尺度上探测样品，获取从纳秒到毫秒的动力学信息，同时在结晶过程中以分钟级的动力学分辨率获取数据。从单体或小聚集体的初始状态到出现大微晶，这个过程需要几个小时。利用中子光谱学使我们能够区分不同的结晶途径，并揭示整个结晶过程中共存聚集体的存在。我们展示了在LaCl存在下从水溶液中结晶的人血清白蛋白（HSA）蛋白质上的多技术方法。对于这个系统，所有方法的结晶动力学都可以用一个S形函数一致地描述，并且动力学可以由盐浓度控制。此外，我们将HSA-LaCl模型系统与β-乳球蛋白-CdCl的结晶行为进行了比较，后者包括一个亚稳态中间状态。
