On the question of two-step nucleation in protein crystallization
Andrea Sauter,Felix Roosen-Runge,Fajun Zhang,Gudrun Lotze,Artem Feoktystov,Robert M. J. Jacobs,Frank Schreiber +6 more
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TLDR
This real-time study provides evidence for a two-step nucleation process for protein crystallization, but also elucidates the role and the structural signature of the MIPs in the nonclassical process ofprotein crystallization.Abstract:
We report a real-time study on protein crystallization in the presence of multivalent salts using small angle X-ray scattering (SAXS) and optical microscopy, focusing particularly on the nucleation mechanism as well as on the role of the metastable intermediate phase (MIP). Using bovine beta-lactoglobulin as a model system in the presence of the divalent salt CdCl2, we have monitored the early stage of crystallization kinetics which demonstrates a two-step nucleation mechanism: protein aggregates form a MIP, which is followed by the nucleation of crystals within the MIP. Here we focus on characterizing and tuning the structure of the MIP using salt and the related effects on the two-step nucleation kinetics. The results suggest that increasing the salt concentration near the transition zone pseudo-c** enhances the energy barrier for both MIPs and crystal nucleation, leading to slow growth. The structural evolution of the MIP and its effect on subsequent nucleation is discussed based on the growth kinetics. The observed kinetics can be well described, using a rate-equation model based on a clear physical two-step picture. This real-time study not only provides evidence for a two-step nucleation process for protein crystallization, but also elucidates the role and the structural signature of the MIPs in the nonclassical process of protein crystallization.read more
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A Review of Classical and Nonclassical Nucleation Theories
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References
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Journal ArticleDOI
Stable prenucleation calcium carbonate clusters.
TL;DR: It is demonstrated that dissolved calcium carbonate in fact contains stable prenucleation ion clusters forming even in undersaturated solution, and specified complex crystal structures challenge the classical view on nucleation considering the formation of metastable ion clusters.
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Enhancement of Protein Crystal Nucleation by Critical Density Fluctuations
TL;DR: Because the location of the metastable critical point can be controlled by changing the composition of the solvent, the present work suggests a systematic approach to promote protein crystallization.
Journal ArticleDOI
Pre-nucleation clusters as solute precursors in crystallisation
TL;DR: In this review, classical nucleation theory, as well as established concepts of spinodal decomposition and liquid-liquid demixing, is introduced together with a description of the recently proposed pre-nucleation cluster pathway.